1. No category

European Conference
of National Strategies for Chlamydia
Trachomatis and Human Papillomavirus
May 25-27, 2011, Jurmala, Latvia
Baltic Beach hotel
CONFERENCE BOOK
Please visit Conference webpage: www.cthpv.org
Auspices
Conference arises from the Project European
Conference of National Strategies for Chlamydia
Trachomatis and Human Papillomavirus-NSCP which
has received funding from the European Union, in
the framework of the Public Health Programme.
1
cobas 4800 CT/NG Test
cobas
CT/NG
Test
Robust4800
CT and
NG testing
in a simplified workflow
®
®
Robust CT and NG testing in a simplified workflow
Multiple Testing Options
This test is being designed to run as CT only, NG only, or as
CT/NG combination.
Multiple Testing Options
This test is being designed to run as CT only, NG only, or as
Plans include
validating test for use at launch with endocervical swabs
CT/NG
combination.
Multiple Specimen Types
Plans include validating test for use at launch with endocervical swabs
as well as male and female urine. Other
specimen types or collection
Roche proprietary collection kit cobas® PCR media to ensure
devices
must
beroom
validated
by customers.
at least 90
days
temperature
sample stability for urine and swab
Multiple Specimen Types
as well as male and female urine. Other specimen types or collection
devices must be validated by customers.
New Collection Kit for Longer
Sample Stability & Primary Tube
samples. The cobas® PCR media tubes can be automatically loaded
Loading onto the Automated
®
on to the
system without
transfer
secondary
Preparation
PCRtube.
media to ensure
Roche
proprietary
collection
kit to
cobas
NewSample
Collection
Kit for Instrument
Longer
Sample Stability & Primary Tube at least 90 days room temperature sample stability for urine and swab
®
•400
µL for
swab
samples
10% of the available
samples.
The
cobas
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mediaequals
tubesless
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be automatically
loaded
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Volume
Required Instrument
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Minimal Sample Input
Volume Required
sample
cobas® PCR
media
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µL collected
for swabinsamples
which
equals less than 10% of the available
®
PCRand
media
sample
cobas
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specimenincan
be stored
used for repeat testing
if necessary
•850
µL for urine samples which equals less than 10% of the available
sample collected in cobas® PCR media
Higher Sensitivity for Detecting
CT & NG with Automated Front
End Sample Preparation
Higher Sensitivity for Detecting
CT & NG with Automated Front
End Sample Preparation
2
Sample preparation is based on state-of-the-art magnetic glass
Residual
specimenThis
canisbesuperior
stored to
and
used
for repeat
testing
particles chemistry.
crude
process
methods
by
ifremoving
necessary
inhibitory materials and concentrating target DNA, resulting
in a more robust and reliable test.
Sample preparation is based on state-of-the-art magnetic glass
particles chemistry. This is superior to crude process methods by
removing inhibitory materials and concentrating target DNA, resulting
in a more robust and reliable test.
High Throughput
The cobas® 4800 System is designed to process up to 384 samples
per day.
Less Hands-On Time
Primary collection tubes can be placed directly onto the sample
preparation instrument. Automation eliminates the need for manual
handling of samples, reducing time and the potential for cross
contamination. Manual processing intervention is limited to the transfer
of the plate to the cobas z 480 analyzer.
High NG Specificity
Newly designed test that targets a well conserved region (DR-9) along
with a specific variant sequence unique to the Neisseria gonorrhoeae
species. The NG assay includes a total of 4 primers and 2 probes
which amplify and detect both wild type and variant versions of this
target sequence.
No cross reactivity with commensal Neisseria or other bacterial species
have been observed with this new NG assay.
Dual target for
C. trachomatis detection
The cobas® 4800 CT/NG Test simultaneously detects two DNA targets;
one within the C. trachomatis cryptic plasmid and the other in the CT
genome. Therefore, targeting infections caused by wild type CT, the
Swedish variant, or other chlamydia strains that may harbor deletions in
the cryptic plasmid, as well as CT that do not carry the cryptic plasmid.
Robust Combo Test
Roche tests are designed to be robust and minimize the effects of PCR
competition in a multi-target assay.
The cobas® 4800 CT/NG Test is being designed to detect CT or NG
targets within a 1:10,000 ratio of each other (10 copies of CT in the
presence of 100,000 copies of NG; 10 copies of NG in the presence
of 100,000 copies of CT).
Quality Controls
•Internal Control for full process monitoring – from extraction
to detection, ensuring effective sample preparation, amplification,
and target detection
•Positive and Negative Controls are included in every run –
to validate the results obtained
•AmpErase enzymes controls contamination from amplicon carry over
Clear and Definitive Results
Results can be Positive, Negative, or Invalid.*
No indeterminate results are expected; reduces need for retesting.
* Invalid results identify specimen inhibition detected by the Internal Control
as means to prevent false negative results.
Workflow Timeline
Total run time to result:
Sample prep:
Detection:
Hands-on time:
4.5 hours per run – 94 patient samples
2 hours
2 hours
30 minutes (estimated)*
Next run can start as soon as the PCR plate is transferred from
cobas x 480 instrument. No centrifugation or incubation required.
* Limited to loading of samples and reagents and transfer of microwell plate
from cobas x 480 instrument to cobas z 480 analyzer.
Expected Performance:
Analytical Sensitivity
Design goal for Limit of Detection is one (1) Inclusion-Forming Unit
(IFU) per PCR for all serotypes of Chlamydia trachomatis.
Design goal for Limit of Detection is ten (10) Colony-Forming Units
(CFU) per PCR for all genotypes of Neisseria gonorrhoeae.
1
INVITATION . ........................................................................................... 3
ORGANIZERS . ........................................................................................ 4
REGISTRATION . ..................................................................................... 5
SCIENTIFIC AND SOCIAL PROGRAMME . ......................................... 8
SUBMITTED ABSTRACTS ................................................................... 12
2
INVITATION
You are very welcome to this Conference arises from the
Project European Conference of National Strategies for
Chlamydia Trachomatis and Human Papillomavirus-NSCP
which has received funding from the European Union, in
the framework of the Public Health Programme.
Target group of the participants includes laboratory
specialists, clinicians, microbiologists, surveillance
experts, public health specialists, healthcare economists,
healthcare funders, policy makers.
Objective of the Conference is evaluation and
developing of Chlamydia trachomatis and HPV current
control programs. Control programs aim to reduce the
prevalence diseases, but this is difficult to monitor as it
requires periodic population surveys of the population.
However, there are many other indicators of the
effectiveness which should be built into any program
from the outset. At the national level, programs should
monitor indicators relating to the policies and guidelines
of the program, the implementation and processes,
and the outcome of the program. These must be based
on the specific objectives appropriate to the level of
implementation. If countries move from one level of
control to the next, they will need to make decisions
based on a rigorous appraisal of the evidence for
effectiveness, cost-effectiveness and harms. This will be
assisted if countries ensure that all activities are fully
evaluated and results shared with others in Europe.
This way investments in programs made now will
strengthen the evidence base for Chlamydia trachomatis
and HPV control and facilitate future decision making
and improve population health. At the European level,
the objective should be to reduce the proportion of
countries reporting no organized activity.
Prof. Aija Zilevica, Lead microbiologist
from the University of Latvia,
Conference President
Prof. Mario Milco D’Elios,
Lead immunologist from the
University of Florence, Chairperson of
the Conference Scientific Committee
Dzintars Ozolins, MD, PhD
President of the Latvian Society
of Laboratory specialists,
Chairperson of the Conference
Steering Committee
INVITATION
Purpose of the Conference is to provide guidance
in the European Union about national strategies for
Chlamydia trachomatis and human papillomavirus (HPV)
early detection and control. It will provide a framework
for developing, implementing or improving national
strategies to control of Chlamydia trachomatis and HPV.
Health policies, like clinical guidelines, should be based
on the best available evidence. In this Conference we
will aim to facilitate the development of local, evidencebased guidelines within the context of sound national
strategies. Such strategies need to take account not
only of clinical and epidemiological factors (such as the
prevalence in the population) but also of local systems
of health care delivery, infrastructure and resourcing.
3
ORGANIZERS
Conference Scientific committee:
Christopher Barbara (Malta), Bertille de Barbeyrac (France), Maria Jose Borrego (Portugal),
Tania Crucitti (Belgium), Alexandros Daponte (Greece), Marius Domeika (Sweden),
Mario Milco D’Elios (Chairperson, Italy), Viorica Gheorghiu (Romania),
Karin Haar (Germany), Bjorn Herrmann (Sweden), Steen Hoffmann (Denmark),
Kai Joers (Estonia), Margaretha Jurstrand (Sweden), Hilde Klovstad (Norway),
Juta Kroica (Latvia), Vesta Kucinskiene (Lithuania), Catherine M Lowndes (Health
Protection Agency), Servaas A. Morre (The Netherlands), Sander Ouburg (The Netherlands),
Dzintars Ozolins (Latvia), Michael Pfleiderer (Germany), Mirja Puolakkainen (Finland),
Dace Rezeberga (Latvia), Magdalena Rosinska (Poland), Peter Sasieni (United Kingdom),
Emma Savage (Health Protection Agency), Par Sparen (Sweden), Magnus Unemo (Sweden),
Inga Velicko (Sweden), Aija Zilevica (Latvia)
Expert group: Mario Milco D’Elios (Chairperson, Italy), Marita van de Laar (ECDC),
Catherine M Lowndes (Health Protection Agency), Dzintars Ozolins (Latvia),
Magnus Unemo (Sweden)
Impact and expected outcomes of the Conference will be realized by Scientific committee
reviewing submitted abstracts for publishing, and providing ideas during meetings of
the Conference for the Expert group. Expert group will ensure scientific control and
long term impact of the Conference results realizing by publishing Conference book
and Essentials of the Conference, preparing questionnaires and summarizing results
from questionnaires. Scientific Committee will confirm abstracts for publishing and their
usefulness for work of the committee during the Conference. This will provide scientific
control of the Conference. Questions arising at the online based Forum for discussion will
be forwarded to Expert group and this will ensure long term monitoring of the impact of
the Conference results.
Conference Steering committee:
Parsla Gredzena, Violeta Mavcutko, Dzintars Ozolins (Chairperson), Vita Scepetova
ORGANIZERS
Steering committee is responsible for organization, planning and management of the
Conference.
4
REGISTRATION
Registration fees include the entrance to the Conference sessions, Conference materials,
coffee brakes, lunch and Gala dinner. Registration fees do not include accommodation
costs and travel expenses.
Registration for accompanying person - entrance to Gala dinner - 60 EUR
REGISTRATION
Registration fees later 15.05.2011 just on site in 26.05.2011 - 300 EUR
5
cobas 4800 HPV Test
cobas
4800
HPV Test
Clinically
relevant,
reliable, and accurate
®
®
Clinically relevant, reliable, and accurate
Assay Design and
Sample Options
Assay Design and
Sample Options
Designed as a qualitative single tube multiplex assay that
simultaneously detects 14 high-risk genotypes, identifies
Designed
a 18,
qualitative
single
multiplex
assay
that
HPV Type as
16 &
and ß-globin
is tube
used as
an internal
control.
simultaneously detects 14 high-risk genotypes, identifies
Test options include:
HPV Type 16 & 18, and ß-globin is used as an internal control.
•HPV-HR only (which provides qualitative result)
•HPV-HR
+ Genotype
Test options
include:(GT) 16/18 (which provides HPV-HR result
and separate
qualitative
resultqualitative
for Type 16 result)
and Type 18)
•HPV-HR
only (which
provides
Multiple Specimen Types
Multiple Specimen Types
Primary Vial Loading On the
Automated Sample Preparation
Instrument
•HPV-HR + Genotype (GT) 16/18 (which provides HPV-HR result
®
TM
and
Liquid
Plans
validation
in bothresult
PreservCyt
andinclude
separate
qualitative
for Type
16SurePath
and Type
18)
Based Cytology (LBC) vials. A Roche labeled specimen collection kit
is also planned for stand-alone DNA tests for samples run separately
Plans include validation in both PreservCyt® and SurePathTM Liquid
or cases where conventional PAP smears are common.
Based Cytology (LBC) vials. A Roche labeled specimen collection kit
is also planned for stand-alone DNA tests for samples run separately
Sample carriers are available to load primary LBC vials directly
or cases where conventional PAP smears are common.
onboard the sample prep system, minimizing hands on time for
transfer of sample.
Primary Vial Loading On the
Automated
Preparation
Minimal Sample
Sample Input
Instrument
Volume Required
Sample carriers are available to load primary LBC vials directly
onboard
prep system,
minimizing
on time for
Only 400the
µLsample
input volume
is required
for each hands
test. Depending
on
transfer
sample.
residual of
volume
remaining from PAP process, options include: loading
Minimal Sample Input
Volume Required
Only 400 µL input volume is required for each test. Depending on
residual volume remaining from PAP process, options include: loading
Sample preparation is based on the state-of-the-art magnetic glass
primary
vial or transferring to a secondary tube (requires 800 µL total
particles chemistry. Onboard heating and mixing of media and cervical
volume). Ideal when residual samples are below 4 mL.
State-of-the-Art Sample Prep
Sample
preparation
contamination
risk. is based on the state-of-the-art magnetic glass
particles chemistry. Onboard heating and mixing of media and cervical
®
cells
provides
superior
preparation
product.
Specialized
The cobas
4800
Systemsample
is designed
to process
up to 280
samples
pipetting
technology
combined
with
enzymes
cross
in one day.
One test can
provide up
to 3AmpErase
results: (HPV-HR,
GTreduces
16,
GT 18) to enhance
contamination
risk.testing efficiency.
State-of-the-Art Sample Prep
primary vial or transferring to a secondary tube (requires 800 µL total
volume). Ideal when residual samples are below 4 mL.
cells provides superior sample preparation product. Specialized
pipetting technology combined with AmpErase enzymes reduces cross
High Throughput
High Throughput
The cobas® 4800 System is designed to process up to 280 samples
in one day. One test can provide up to 3 results: (HPV-HR, GT 16,
GT 18) to enhance testing efficiency.
Less Hands-On Time
Collection tubes are placed directly onto the sample preparation
instrument, eliminating the need for manual handling of samples,
reducing time and the potential for cross contamination. Manual
processing intervention is limited to the transfer of the plate to the
cobas z 480 analyzer for amplification and detection.
Clinical Relevance
Newly designed primers and probes target a well defined L1
region for DNA
Expect to correlate to detect medically relevant pre-cancer ≥ CIN2+
Medical decision point to balance clinical specificity and clinical
sensitivity to ensure application is suitable for screening and to achieve
guidelines and benchmarks 1
Information on infection with genotypes 16 and 18, which have been
shown to increase risk for cervical disease and provide “actionable”
data for immediate follow-up 2
Assay designed to correctly identify genotypes important for HPV–HR
panel and not to cross react with low–risk types
Robust PCR Test
Quality Controls
•Internal Control ( ß-globin) for full process monitoring –
from extraction to detection, ensuring effective sample preparation,
amplification, and target detection
•Positive and Negative Controls are included in every run –
to validate the results obtained
•AmpErase enzymes helps control contamination from
amplicon carry over
Results can be Positive, Negative, or Invalid.*
Clear and Definitive Results
No indeterminate results are expected; reduces need for retesting.
* Invalid results identify specimen inhibition detected by the Internal Control
as means to prevent false negative results.
Total run time to result:
Sample prep:
Detection:
Hands-on time:
Workflow Timeline
5 hours per run – 94 patient samples
2 1/2 hours
2 hours
30 minutes (estimated)*
Next run can start as soon as the PCR plate is transferred from
cobas x 480 instrument. No centrifugation or incubation required.
* Limited to loading of samples and reagents and transfer of microwell plate
from cobas x 480 instrument to cobas z 480 analyzer.
1
Meijer Chris JLM, Berkhof, J, Castle PE et al. Guidelines for human papillomavirus DNA test
requirements for primary cervical screening in women over 30 years and older. Int J of Cancer
124, 516-520 (2009)
2
Khan MJ, Castle PE, Lorincz AT et al. Elevated 10 year risk of cervical precancer and cancer in
women with human papillomavirus (HPV) Type 16 or 18 and the possible utility of type specific
HPV testing in clinical practive. J Natl Cancer Inst. 2005:97:1072-1079
ROCHE, COBAS, LIFE NEEDS ANSWERS,
and AMPERASE are trademarks of Roche.
PreservCyt is a registered trademark of Hologic, Inc.
SurePath is a registered trademark of Becton, Dickinson and Company
© 2009 Roche Molecular Systems, Inc.
Roche Molecular Diagnostics
4300 Hacienda Drive
Pleasanton, CA USA 94588
http://molecular.roche.com
SCIENTIFIC AND SOCIAL PROGRAMME
25.05.2011
Workshop of Scientific committee – CLOSED MEETING. Preparing Essentials of the
Conference using Conference book and latest approaches from Expert side
26.05.2011
9:00 - 12:00; 10:30 - 11:00 Coffee break
Plenary session (Baltic Beach hotel, room Jura + Jurmala). Survey of Chlamydia
surveillance in Europe. Leading specialists are invited to present current surveillance
systems in different European countries focusing on strengths as well as weaknesses.
Chairpersons: Aija Zilevica (Latvia), Charlie Jennison (Health Protection Agency),
Karin Haar (Germany)
9:00 - 9:10 Opening. Aija Zilevica (Latvia)
9:10 - 9:40 Charlie Jennison (Health Protection Agency) “Surveillance of Chlamydia
trachomatis in England: a regional perspective”
9:40 - 9:55 Questions, discussion
9:55 -10:15 Karin Haar (Germany) “Epidemiology of Chlamydia trachomatis infections in
Germany: Data from the STD-Sentinel”
10:15-10:30 Questions, discussion
11:00-11:20 Steen Hoffmann (Denmark) “Laboratory Surveillance of Chlamydia
trachomatis in Denmark 1994 - 2010”
11:20-11:40 Jurijs Perevoscikovs (Latvia) „Epidemiological surveillance of Chlamydia
trachomatis infection in Latvia”
11:40-12:00 Questions, discussion
Satellite session (Baltic Beach hotel, room Krasts I+II). Genotyping of Chlamydia
trachomatis. Chairpersons: Margaretha Jurstrand (Sweden), Bjorn Herrmann (Sweden)
PROGRAMME
9:00 - 9:20 Mirja Puolakkainen (Finland) “Genotyping of Chlamydia trachomatis strains”
9:20 - 9:50 Margaretha Jurstrand (Sweden) “Detection of Chlamydia trachomatis by
genetic methods in a Swedish county before and after identification of the
new mutated variant”
9:50 -10:20 Bjorn Herrmann (Sweden) “Epidemiological knowledge by genotyping
Chlamydia trachomatis: an overview of recent achievements”
10:20-10:30 Questions, discussion
11:00-11:30 Magnus Unemo (Sweden) “The amazing story of the Swedish new variant of
Chlamydia trachomatis”
8
11:30-11:50 Kai Joers (Estonia) “Prevalence and genetic structure of Chlamydia
trachomatis population in Estonia”
11:50-12:00 Questions, discussion
Satellite session (Baltic Beach hotel, room Selga). Routine diagnostics of Chlamydia
and HPV. What about cost-efficiency? Chairpersons: Jolande Land (The Netherlands),
Tania Crucitti (Belgium)
9:00 - 9:30 Jolande Land (The Netherlands) “Chlamydia antibodies as markers for late
complications in cost-effectiveness models”
9:30 - 9:50 Alexandros Daponte (Greece) “Self sampling and HPV DNA test. What is the
evidence?”
9:50 -10:20 Tania Crucitti (Belgium) “Cost-effective testing for Lymphogranuloma
venereum: the experience in Belgium”
10:20-10:30 Questions, discussion
11:00-11:20 Vesta Kucinskiene (Lithuania) “Pooling genital samples could be a cost beneficial
approach for screening C. trachomatis in young female patients in Lithuania”
11:20-11:30 Questions, discussion
12:00 - 14:00 Lunch
14:00 - 17:00; 15:30 - 16:00 Coffee break
Plenary session (Baltic Beach hotel, room Jura + Jurmala). Screening programmes of
Chlamydia in Europe. The analyzing of the different screening programmes in Europe
will be presented. Focusing on ways how as high response rates as possible shall be
achieved. Chairpersons: Marita van de Laar (ECDC), Per-Anders Mardh (Sweden)
14:00-14:30 Marita van de Laar (ECDC) “Chlamydia control in the Europe”
14:30-15:00 Inga Velicko (Sweden) “Epidemiology and control of genital Chlamydia
trachomatis infection in Sweden”
PROGRAMME
15:00-15:30 Sebastian Kalwij (United Kingdom) “8 years of Chlamydia screening in
England, experiences and lessons learnt”
16:00-16:30 Per-Anders Mardh (Sweden) “Epidemiological aspects on surveillance and
screening”
16:30-16:50 Jorma Paavonen (Finland) “Prevention of C.trachomatis and HPV infections:
Are we doing enough?”
16:50-17:00 Questions, discussion
9
Satellite session (Baltic Beach hotel, room Krasts I+II). HPV surveillance and vaccine era.
Chairpersons: Michael Pfleiderer (Germany), Emma Savage (Health Protection Agency)
14:00-14:30 Emma Savage (Health Protection Agency) “HPV surveillance in the vaccine era”
14:30-14:50 Peter Sasieni (United Kingdom) “How to set up a cervical screening
programme if none exists”
14:50-15:20 Michael Pfleiderer (Germany) “Current understanding of safety and efficacy
of HPV vaccines licensed in the EU”
15:20-15:30 Questions, discussion
16:00-16:20 Viorica Gheorghiu (Romania) “Challanges and facts on HPV vaccination in
Romania”
16:20-16:30 Questions, discussion
19:00-22:00 Social programme (Gala dinner in Baltic Beach hotel 1st floor restaurant Perle and Caviar club). Algirdas Suminskas and group
“Keksi” performance
27.05.2011
10:50-11:20 Coffee break
9:00-10:50
Plenary session (Baltic Beach hotel, room Jura + Jurmala). National laboratory
diagnostics programmes of Chlamydia trachomatis. Presentations are focused on
suggestions how the best practices shall be implemented in every European country.
Chairpersons: Mario Milco D’Elios (Italy), Marius Domeika (Sweden)
9:00- 9:30 Max Chernesky (Canada) „Screening Strategies for Chlamydia and High Risk
HPV: What are the Roles for Self Collection and Amplified DNA and RNA
Assays?”
PROGRAMME
9:30- 9:50 Marius Domeika (Sweden) “Quality enhancements and quality assurance of
laboratory diagnosis of sexually transmitted infections in Eastern Europe”
9:50-10:00 Questions, discussion
10:00-10:20 Diana Dusacka (Latvia) “Detection of Chlamydia trachomatis by molecular
biological methods in Latvia”
10:20-10:30 Questions, discussion
10:30-10:40 Mario Milco D’Elios (Italy) “Actual issues from the Conference Scientific
committee”
10:40-10:50 Closing remarks. Dzintars Ozolins (Latvia)
10
9:00-10:50
Sattelite session (Baltic Beach hotel, room Krasts I+II). Advances in the diagnosis
of Chlamydia and HPV. Scientifically oriented session concerning development
of the new laboratory methods. Chairpersons: Sven Müller-Loennies (Germany),
Sander Ouburg (The Netherlands)
9:00 - 9:30 Edwin Roovers (The Netherlands) “Do Abbott RealTime CT and High Risk
HPV meet the requirements for laboratory diagnosis and screening of CT
and HPV?”; sponsored by Abbott Molecular
9:30 - 9:50 Sander Ouburg (The Netherlands) “The value of host genetic markers
combined with CT serology in tubal pathology diagnosis”
9:50 -10:20 Frank Pieksma (Switzerland) “Cobas 4800: new developments for the
detection of HPV & CT/NG”; sponsored by Roche
10:20-10:30 Questions, discussion
10:30-10:50 Sven Müller-Loennies (Germany) “Antibodies against Chlamydia
Lipopolysaccharide -- Molecular Insight into Specificities and CrossReactions”
9:00-10:50
Round table (Baltic Beach hotel, room Selga). New approaches to manage
Chlamydia and HPV screening. Chairpersons: Bertille de Barbeyeac (France),
Catherine M Lowndes (Health Protection Agency)
9:00 - 9:20 Jolita Rimiene (Lithuania) “New methods for cervical cancer detection in
Lithuania. Value of high-risk HPV-DNA testing in the triage of women with
ASC category from liquid based cytology”
9:20 - 9:40 Maria José Borrego (Portugal) “Lymphogranuloma venerum recent outbreak
in Europe: the Portuguese experience”
9:40 - 9:50 Questions, discussion
9:50 -10:20 Jean-Jacques Palombo (Switzerland, Roche Scientific Division) “Advances in
the diagnosis of Chlamydia and HPV - what happens in industry?”
PROGRAMME
10:20-10:40 Bertille de Barbeyrac (France) “Is glans swab appropriate for diagnosis of
Chlamydia trachomatis infection in asymptomatic men?“
10:40-10:50 Questions, discussion
Commercial presentations and/or fairs
Roche - Baltic Beach hotel, room Dzintars
Abbott Molecular - Baltic Beach hotel, room Liedags
Gen-Probe - Baltic Beach hotel, room Banga
BD - Baltic Beach hotel, room Krasts III
11
SUBMITTED ABSTRACTS
IS GLANS SWAB APPROPRIATE FOR DIAGNOSIS OF C.TRACHOMATIS INFECTION IN
ASYMPTOMATIC MEN?
Corresponding author: Bertille de Barbeyrac
Institution of corresponding author: Université de Bordeaux, INRA, USC Mycoplasmal and
chlamydial infections in humans, French National Reference Center for chlamydial infections,
33076 Bordeaux, France
Country: France
E-mail: Bertille.de.Barbeyrac@u-bordeaux2.fr
With nucleic acid amplified tests (NAATs) that are sensitive and specific for detection of
C. trachomatis, non-invasive specimens such as first catch urine (FCU) and self-collected
vaginal swabs can be used. While FCU is the alternative to the urethral swabs, specimen
transport can be messy and problems have been associated with the handling of FCU in the
laboratory. Swabs are a better specimen for processing. A few studies have shown that
self-collected glans swab may be appropriate for C. trachomatis NAATS. We evaluated
self-collected glans swab vs FCU in asymptomatic heterosexual men and MSM attending
the screening center (anonymous and free of charge) in Bordeaux, France [1].
A subject first self-collected penile swab using a flocked regular swab (Copan Italia Spa, Italy,
Mast Diagnostic, France) and after self-collected urine. The flocked swab was transported
dry to the laboratory and transferred into M4RT transport medium (500µl) (method 1) or
into urine (500µl) (method2). Both samples were simultaneously tested for C. trachomatis
using the PCR assay (Cobas TaqMan CT Test, Roche Diagnostic). The load of C. trachomatis
was estimated by using cycle thresholds (Cts values) given by Cobas TaqMan in all positive
specimens.
ABSTRACTS
For 344 men included in method1, the sensitivity was 89% (24/27) for urines and 67%
(18/27) for penile swabs. The comparison of means of Cts values showed a difference of at
least 4 Cts between urine (33.2) and penile swab (38.5) when both were positive or when
only one was positive, urine (38) and penile swab (42). For 259 men included in method
2 (pooling strategy), the sensitivity of 94.7% was identical for urines and for penile swabs
discharged in urine. The comparison of means of Cts values showed similar Cts between
urine (35.4) and penile swab in urine (36.7). When only one test was positive, Cts were high,
41.2 for urines and 40.7 for penile swabs.
In asymptomatic men, the number of bacteria is often very low and could explain –
the discrepancies of both specimens, - the low sensitivity of penile swab, - the non
reproducibility of results. Our results agree with those of two others studies [2, 3]. These
results suggest the self-collected penile swab may not be useful for C. trachomatis testing.
The penile swab could not replace FVU in screening programs
1 Raherison S, Peuchant O, Clerc M, et al. Glans swabs are not appropriate specimens for
diagnosis of Chlamydia trachomatis infection in asymptomatic men. J Clin Microbiol. 2009;
47: 2686.
12
2 Moncada J, Schachter J, Liska S, Shayevich C, Klausner JD. Evaluation of self-collected
glans and rectal swabs from men who have sex with men for detection of Chlamydia
trachomatis and neisseria gonorrhoeae by use of nucleic acid amplification tests.
J Clin Microbiol. 2009; 47: 1657-1662.
3 Pittaras TE, Papaparaskevas J, Houhoula DP, et al. Comparison of penile skin swab with
intra-urethral swab and first void urine for polymerase chain reaction-based diagnosis of
Chlamydia trachomatis urethritis in male patients. Sex Transm Dis. 2008; 35: 999-1001.
LYMPHOGRANULOMA VENEREUM RECENT OUTBREAK IN EUROPE: THE
PORTUGUESE EXPERIENCE
Corresponding author: Maria José Borrego
Other authors: Gomes JP* Nunes A* Florindo C* Borges V* Ferreira R* Santo I** Azevedo J**
Institution of corresponding author: National Institute of Health, Lisbon, Portugal
Institutions of other authors: *National Institute of Health, Portugal **STD clinic,
Lapa Health Center, Lisbon, Portugal
Speciality: Molecular microbiology
Country: Portugal
E-mail: M.Jose.Borrego@insa.min-saude.pt
Introduction: In Portugal no surveillance measures were launched as a consequence of
the 2004 lympgogranuloma venereum (LGV) outbreak in Europe. However, all Chlamydia
trachomatis specimens that are sent to the National Institute of Health (NIH) of Portugal are
systematically genotyped. This procedure potentially enables the opportunistic detection of
Portuguese circulating LGV strains.
Objectives: To report the unusual number of LGV specimens observed during the 20072010 period, and to analyze clinical and epidemiologic data of LGV infected individuals.
Additionally, to compare the outer membrane protein A gene (ompA) sequence of LGV
specimens with both L2-434 reference strain and L2-variants described in the literature.
Results: So far, a total of 514/833 (61,7%) C. trachomatis specimens could be ompA
genotyped and 17/514 (3,3%) matched LGV-genotypes (16 "L2" and 1 mixed E+L2
undetermined variant). LGV-genotypes were observed in both men (n=10; 5 MSM+1
heterosexual+4 unknown; 4 presented proctitis of which 2 were HIV+) and women (n=7;
all heterosexual, 6 asymptomatic and 1 PID). All the LGV samples detected in 2007 revealed
ompA sequences different from both L2-434, and L2b-144276. LGV specimens detected
during 2008 and 2010 were similar to L2b-144276, while the ones detected during 2009
were similar to the L2 prototype strain.
13
ABSTRACTS
Methods: During the 2007-2010 period, C. trachomatis infection was tested over 9000
samples by the Cobas-Amplicor (2007-2009) or the Cobas 4800 (2010), yielding 833 (9,2%)
positive results. DNA was extracted from C. trachomatis-positive samples, using the
QIAamp DNA Mini Kit. Amplification and automated-sequencing of the ompA gene, using
Laser-Gene99 software (DNASTAR), were used for genotyping.
Discussion: From 1991 to 2002 only 5/463 typed specimens were L2 strains. During 2002-2006
no LGV strain was detected. So, the recent number of L2 specimens was quite unexpected,
probably related with the recent worldwide outbreak of LGV. During 2007 two studies
(1 Portuguese, 1 Austrian) reported new L2 variants, but no link could be established with
specific clinical findings. Globally, the majority of the amino acid changes on the L2 variants
occur in a well-defined antigenic domain.
Conclusions: The particular location of the mutations of the newly described L2 variants
suggests a chlamydial strategy for host immune evasion. The lack of surveillance systems
for C. trachomatis infections, including LGV, in Portugal, makes this outbreak has passed
completely unnoticed and prevented the taking of any control measures or to establish any
link between the Portuguese LGV cases and European (or other) sexual networks.
KNOWLEDGE, ATTITUDES AND BEHAVIOUR ABOUT SEXUALLY TRANSMITTED
DISEASES (STDs): A SURVEY AMONG ITALIAN UNIVERSITY FEMALE STUDENTS
Corresponding author: Chiara Cadeddu
Other authors: Chiara de Waure*, Maria Rosaria Gualano*, Alice Mannocci°,
Giacomina Chiaradia^, Daniela Vincitorio°°, Francesco di Stanislao°°, Elisabetta De Vito**,
Elisa Langiano**, Antonio Boccia°, Walter Ricciardi*, Giuseppe La Torre°
Institution of corresponding author: Hygiene Institute - Catholic University of Sacred Heart
(Rome, Italy)
Institutions of other authors:
*Institute of Hygiene, Catholic University of the Sacred Heart, Rome, Italy
°Clinical Medicine and Public Health Unit, Sapienza University of Rome, Rome, Italy
^Spallanzani Institute, Rome, Italy
°°Section of Hygiene and Public Health, Polytechnic University of the Marche Region, Italy
**Department of Motor Sciences and Health, Local Health Agency, Cassino, Italy
Speciality: Hygiene & Preventive Medicine
Country: Italy
ABSTRACTS
E-mail: chiaracadeddu@yahoo.it
Background: Sexually Transmitted Diseases (STDs) represent a considerable Public Health
issue. Although in the second half of the last century the diffusion of effective screening and
prevention tools has made possible important improvements in terms of mortality, there
is still a great concern about STDs and young women appear as the most affected by the
“development deficit” of the potential of STDs prevention.
Objectives: To find out the level of knowledge and attitudes about STDs and preventive
measures among young women and deepen the understanding of variables associated to
them.
14
Methods: We designed a questionnaire accounting mostly for knowledge about STDs
and knowledge and attitudes toward Pap test and vaccination against Human Papilloma
Virus (HPV), and submitted it to 313 young female university students (mean age: 22 ys),
opportunistically identified. Chi-square test and Mann-Whitney test were performed in order
to identify associated factors.
Results: Knowledge about STDs seems to be weak, in contrast to knowledge about
preventive measures, which appears good but limited. About STDs ways of transmission,
85.3% of women who answered that the use of condom and intercourses with a single
known partner are effective behaviours for preventing STDs showed a right knowledge
of STDs ways of transmission in comparison to 67.2% of women giving incorrect answers
(p<0.001). Eighty three point two per cent of women who took information about STDs
from Internet, books and TV showed a correct knowledge of STDs ways of transmission, in
comparison to 67.3% (p=0.002) among the remainder.
For Pap test the following results came up: 100% of women with a high level of knowledge
on STDs knew Pap test in contrast to less than 90% among the remainder (p=0.04); moreover
89.4% of the women that were aware of effective behaviours for preventing STDs (condom
and intercourses with a single known partner) knew the Pap test in comparison to 80.8% of
the women that were unaware of them (p=0.04).
Conclusions: Women are still not completely aware of risks for STDs and preventive measures
to avoid them. Informational and educational campaigns should be organised to reach this
target and limit the current and future burden of STDs.
REVIEW OF GENITAL CHLAMYDIOSIS AND RISK FACTORS IN LITHUANIA
IN 2007 - 2009
Corresponding author: Caplinskiene Irma
Institution of corresponding author: Lithuanian Centre for Communicable Diseases and AIDS,
Mykolas Romeris University
Speciality: Medical epidemiologist, MD
Country: Lithuania
Introduction: The incidence of Chlamydia trachomatis in Lithuania was to 9.6 cases per
100000 persons in 2009, and in comparison with 2006 (11.9 cases per 100000 persons) it has
decreased. There were 326 cases of Chlamydia infection registered in 2009, and 403 cases in
2007 and 2008 in Lithuania. Though, it is believed that the incidence of Chlamydia infection
is substantially higher than that officially recorded.
Objectives: To assess the trends of Chlamydia trachomatis incidence and related risk factors
(behavioral and sociodemographic) over the last 3 years (2007-2009) in Lithuania.
Methods: Statistical analysis of data (sociodemographic characteristics and sexual risk
behavioral factors) of Chlamydia trachomatis infection cases through the period of
2007-2009. The data was accumulated by the National Centre for Communicable
Diseases and AIDS.
15
ABSTRACTS
E-mail: irma@ulac.lt
Results: Trends: steady increase of chlamydia cases among 20-29 year old male who had 1-3
female sex partners; the number of chlamydia cases among those who indicated a casual
partner as a likely source of infection increased; however, the number of chlamydia cases
among those who could not tell the likely source of infection decreased. There were fewer
cases of infection among individuals who reported constant use of condoms, whereas
the number of chlamydia cases among those who reported occasional use or non-use of
condoms increased; no differences were found in terms of education, place of residence, age
group, social insurance, social groups, and disease detection place, compared to 2007, 2008
and 2009.
Conclusions: There is an annual increase of genital chlamydia infections in the group of
young males from urban areas. There were fewer cases among individuals who reported
constant use of condoms. No differences were found in terms of education, place of
residence, age group, social insurance, social groups, disease detection place, and whether
or not the person was engaged in a commercial sex scenario, compared to 2007, 2008 and
2009. The contact tracing deteriorates with each year.
SCREENING STRATEGIES FOR CHLAMYDIA AND HIGH RISK HPV: WHAT ARE THE ROLES
FOR SELF-COLLECTION AND DNA AND RNA ASSAYS?
Corresponding author: Max Chernesky, PhD
Institution of corresponding author: McMaster University/St. Joseph’s Healthcare, Hamilton,
ON, Canada
Country: Canada
Introduction: Sexually transmitted Chlamydia trachomatis (CT) and human papillomaviruses
(HPV) are very common worldwide. Because the majority of these infections are
asymptomatic, screening programmes using non-invasive sampling methods are required
to identify and treat lower genital tract CT infections to prevent ascending upper tract
complications. The screening goal for HPV testing is to test women for high risk HPV
adjunctively to cervical Pap testing or vaginal testing of women who decline pelvic
examination, in order to identify and treat women with precancerous lesions. Sensitive and
specific nucleic acid assays are available for these purposes.
ABSTRACTS
Objectives: To review comparative data on the use of commercial assays for CT and high risk
(HR) HPV using cervical swabs and Pap collections versus vaginal and urine specimens in
women and urethral, urine, penile meatal, anal and oral swabs in men.
Methods: Publications from the peer-reviewed literature and recent data from the author’s
laboratory will be presented to provide evidence for using various specimen types and DNA
and rRNA assays for CT diagnosis. The performance of DNA and E6/E7 mRNA assays will be
reviewed for HR HPV diagnosis.
Results: CT infections are most prevalent in sexually active, younger women and men.
Lower genital tract asymptomatic infection rates can be as high as 75% allowing inapparent
ascending infections in women and unknowing transmission of infection between sexual
partners. Specimens of choice for screening are self-collected vaginal swabs for women,
and first catch urine and/or self-collected penile swabs for men. There is good evidence for
16
screening self-collected anal swabs from men who have sex with men. Although women
over 30 are primary candidates for Pap and HPV testing, in some societies Pap tests are
being performed on younger women. These specimens can serve as samples of convenience
for screening for CT. Several commercial molecular tests for CT are being used and in
comparative studies the results from these assays may be impacted by analytical sensitivity,
stability of nucleic acids in specimens, and inhibitors. Comparisons of different collection
devices and manipulation of specimens in the preanalytical phase of testing may influence
testing outcomes. Similar studies for HPV have shown that most specimen types contain
DNA and E6/E7 mRNA. Most of the commercial assays have comparative sensitivities for
detecting CIN2+ pathology and have been shown to facilitate colposcopy examinations for
patients with ASC-US cytology. The role for HPV adjunctive testing and screening in select
populations is becoming clearer.
Discussion: Sampling, testing, automation and focused screening will be discussed with an
emphasis on research gaps.
Conclusions: Manufacturers of clinically relevant diagnostic assays for CT and HPV deserve
credit for advancing our knowledge of the role of these pathogens in patient morbidity.
Screening programmes should use these assays wisely.
CHLAMYDIA SCREENING IN IRELAND - NOT READY YET? RESULTS OF A PILOT
STUDY OF OPPORTUNISTIC SCREENING FOR GENITAL CHLAMYDIA TRACHOMATIS
INFECTION IN THE REPUBLIC OF IRELAND
Corresponding author: Dr. Emer O' Connell 1
Other authors: D Vaughan 2, R Brugha 3, M Balfe 3, M Cormican 2,4, P Gillespie 2, C O' Neill 2,
C Coleman 4, C Fleming 2,4, M Fitzgerald 1, A Murphy 2, D O'Donovan 2,5.
Institution of corresponding author: 1Health Service Executive (HSE) Dublin/Mid-Leinster,
Ireland
Institutions of other authors: 1 Health Service Executive (HSE) Dublin/Mid-Leinster, Ireland 2 National University Ireland, Galway, Ireland 3 Royal College of Surgeons, Dublin, Ireland 4 Galway University Hospitals, Ireland 5 Health Service Executive (HSE) West, Galway, Ireland
Speciality: Public Health Medicine
Country: Republic of Ireland
Introduction: Chlamydia trachomatis (CT) is an infection of public health concern in the
Republic of Ireland with annual notifications rising from 245 in 1995 to 1,278 in 2003. In 2004,
legislation requiring laboratory notification came into effect, and the number increased
to 6,290 in 2008 [1]. CT is now the most commonly notified sexually transmitted infection
(STI) in Ireland. Irish prevalence data shows asymptomatic rates of 3.7-11.2 for females and
5.9% for males. There is no formal organized screening programme for CT in the Republic of
Ireland.
Objectives: A chlamydia screening pilot study was conducted in Ireland between 2007 and
2009 to assess the feasibility and cost effectiveness of introducing a screening programme
17
ABSTRACTS
E-mail: emer.oconnell@hse.ie
for CT in the Republic of Ireland. 18-29 years olds were the eligible population. 16 and 17
year olds were excluded because of ethical restrictions under Irish law; and the range 18 to
29 years includes the highest notification group (20-29 years old) in Ireland.
Methods: Opportunistic screening was offered to women and men aged 18-29 years who
attended urban and rural general practices, student health units, and a family planning
clinic in the west of Ireland. Non clinical 'pee-in-a-pot' (PIP) days were organized in
two higher education institutions. Urine specimens were tested with polymerase chain
reaction technology (COBAS TAQMAN). Economical evaluation compared the pilot strategy
of opportunistic screening in terms of costs, including those averted by screening, and
effectiveness.
Results: 998 eligible people were tested: 460 in clinical settings and 538 in non clinical PIP
settings. Screening offer rates were low (1-9%), refusal rates were low, and urine testing was
highly acceptable. 48 people (4.8%, 95% CI: 3.5-6.1) tested CT positive. Management of cases,
partner notification and outcome rates corresponded well with best international practice.
Screening in the combined clinical settings would result in an incremental cost per quality
adjusted life year (QALY) gained of 94,717 EUR, while 'pee-in-a-pot' screening was estimated
to result in 42,967 EUR per QALY gained.
Discussion: A research health adviser was essential for coordinating the care of positive
cases, especially for partner notification. Low offer rates in clinical settings reflected the time
constraints on General Practitioners and practice nurses. Opportunistic screening in the
combined healthcare settings is not cost-effective. Neither screening settings are considered
cost-effective.
Conclusions: We recommend prioritizing primary prevention and the provision of
supports to primary care providers for optimal case diagnosis (where clinically indicated),
management and follow-up. Dedicated regional health advisers could provide support and
advice to clinicians and cases, and coordinate partner notification, testing and treatment.
References: (1) HPSC. The Health Protection Surveillance Centre. Sexually Transmitted
Infections 2008. Annual summary report. 2010.
COST-EFFECTIVE TESTING FOR LYMPHOGRANULOMA VENEREUM:
THE EXPERIENCE IN BELGIUM
Corresponding author: Tania Crucitti
ABSTRACTS
Other authors: Hilde Smet
Institution of corresponding author: Institute of Tropical Medicine
Institutions of other authors: Institute of Tropical Medicine
Speciality: Microbiology
Country: Belgium
E-mail: tcrucitti@itg.be
18
Introduction: Lymphogranuloma venereum is reported in several countries in Europe. It has
been accepted that the infection is confined to men having sex with men usually presenting
symptoms of proctitis. The testing for L genotypes of Chlamydia trachomatis is very often
restricted to this high risk population. Up to date laboratories rely on in-house methods to
identify the L genotypes which can be costly and time consuming depending on the method
used.
Objectives: To apply a nucleic acid amplification testing algorithm to firstly screen specimens
for C. trachomatis and to secondly identify the non-L and L genotypes without increasing
considerably the cost of testing, hands-on time and turnaround time.
Methods: C. trachomatis positive DNA extracts tested with the Abbott CT/NG Real Time PCR
were re-tested using an in-house Real Time-PCR (RT-PCR) assay to determine whether the
C. trachomatis belonged to the L genotypes. The RT-PCR differentiated L from non L
genotypes. As part of the validation of the in-house RT-PCR its performance was compared
to the Restriction Fragment Length Polymorphism (RFLP) assay used so far. The algorithm
was validated on urine, genital, pharyngeal and anal specimens.
Results: The Abbott CT/NG Real Time PCR and in-house RT-PCR showed identical sensitivities
and specificities for genital, anal, pharyngeal and urine specimens. Fifty two positive anal
specimens were tested in the RFLP and RT-PCR assay. Both methods identified 8 non-L,
42 L and 1 mixed non-L and L type infection. One specimen was a non-L type by RT-PCR but
undetermined using RFLP. Since the introduction of the algorithm in September 2010 and
till February 2011, a total of 1124 specimens were tested; 66 were C. trachomatis positive
with 53 non-L and 13 L genotypes. All results were reported within 2 days after specimen
reception. The routine use of the RT-PCR assay increased the cost with only 10%.
Conclusions: We identified L genotypes of C. trachomatis at an affordable cost, within
a reasonable time frame, and in all C. trachomatis positive specimens. By consequence we
are now able to identify LGV infection in the general population or in patients in which LGV
infection was not suspected. At the end the patients will benefit from a more timely and
reliable diagnosis and appropriate management. Finally we will gain insight into the LGV
epidemiology.
19
ABSTRACTS
Discussion: The presented algorithm improved the diagnosis of LGV considerably. The L
genotypes were identified immediately after obtaining a C. trachomatis positive result
and independently from the origin of the specimen and risk behavior of the patient. This
is in contrast with the previously used RFLP method which was performed on batches of
specimens and on only anal specimens collected from MSM. In addition, the hands-on
time for the in-house RT-PCR method was reduced by using the same DNA extract as the
screening assay.
SELF SAMPLING AND HPV DNA TEST.
WHAT IS THE EVIDENCE?
Corresponding author: Alexandros Daponte
Country: Greece
E-mail: dapontea@otenet.gr
Background: During the last decade, increasing efforts have focused on increasing the overall
proportion of women participating in cervical cancer screening procedures, mainly by HPV
detection in self-obtained samples, such as vaginal and urine specimens as reported by
our team previously (Daponte et al. 2003, 2004). HPV screening during antenatal visits may
increase participation, facilitate early detection and be cost-effective for patients and the
health services.
Objectives: A study of screening high-risk (hr) human papillomavirus (HPV) types in first
void urine and vaginal samples in pregnant women attending the antenatal clinic using our
optimized protocol for PCR detection in urine and vaginal samples in order to establish the
feasibility of using these specimens for HPV detection and compare them with non pregnant
women hr HPV incidence rates from the same area in central Greece (University hospital of
Larissa).
Study design: Paired urine and vaginal specimens obtained from 500 pregnant women
attending antenatal clinic were included. Vaginal swabs and urine samples were tested
in all participants for HPV types 16, 18, 31 and 45 after urine microscopy was performed.
Prevalence of HPV was compared with 500 paired urine and vaginal specimens from non
pregnant women collected by midwives in the same area.
ABSTRACTS
Methods: Urinalysis and urine microscopy was performed. QIAamp Viral RNA Mini Kit
(Qiagen Inc., Valencia, CA) was used for DNA extraction. An in house PCR method was used
for amplifying the above hr HPV types. The quality of sample was validated by detection
of the housekeeping beta-globin gene.
Results: The hr HPV detection rate was similar in pregnant and non pregnant women. In the
antenatal patients, 15/500 paired urine/vaginal samples were both positive for the same type
of hrHPV. Only one sample was vaginal positive/urine negative. The urine/vaginal detection
sensitivity was higher in pregnancy because first void urine samples from pregnant women
usually contain two or more epithelial cells per field in urine microscopy increasing HPV
detection as reported by our team before.
Conclusions: Urine and vaginal samples are easily collected during antenatal visits and
may be particularly important in screening young women who are asymptomatic and for
personal or cultural reasons would not participate in a cervical specimen screening program.
This study indicates that urine and vaginal specimens in pregnancy are easily obtainable and
adequate for screening for hr HPV but this should be confirmed in a larger future prospective
trial.
20
CHLAMYDIA IN SWEDEN - KEY FACTORS OF SUCCESSFUL
PREVENTIVE RESPONSE
Corresponding author: Charlotte Deogan
Other authors: Cecilia Moberg, PhD Anna Månsdotter, PhD Lene Lindberg, PhD, Associate
Professor
Institution of corresponding author: Department of Public Health Sciences,
Karolinska Institutet, Stockholm Sweden.
Institutions of other authors: Department of Public Health Sciences, Karolinska Institutet,
Stockholm Sweden
Speciality: Public health
Country: Sweden
E-mail: charlotte.deogan@ki.se
Introduction: After a continuous increase of Chlamydia trachomatis in Sweden, a general
reduction in the incidence was seen in 2009. Incidence and prevention activities varied
largely between geographical regions.
Objectives: The aim of the study was to identify key factors of successful regional prevention
of chlamydia and other sexually transmitted infection (STIs) by a mapping of the preventive
response in seven Swedish counties and a case comparison related to chlamydia incidence.
Methods: A multiple case study was performed including seven Swedish counties based
on data from surveys and structured interviews with key stakeholders, county council
registry data and national surveillance data on chlamydia incidence and testing. Factors
of prevention were rated in a case comparison across counties resulting in a matrix of
prevention factors. Key factors of successful prevention were identified by contrasting high
incidence counties with low incidence counties.
Discussion: Results were found to be compliant with earlier research. Further research and
evaluation is however needed of specific measures to explore what constitutes an effective
prevention mix.
Conclusions: Successful prevention of chlamydia at the regional level requires strong
leadership and coordination of preventive measures that incorporates multiple collaboration
partners and research knowledge. Adequate resource investments in both primary- and
secondary prevention and a broad mix of prevention activities with high testing coverage
targeting risk individuals are key factors in successful preventive health care response.
21
ABSTRACTS
Results: Counties with a reduction in chlamydia incidence apply adequate investments
in STI-prevention, simultaneously implement a broad mix of activities, involve multiple
local agents, include efforts to target risk individuals with testing and counseling and use
the internet as a platform for prevention activities. Other key factors identified were a
strong leadership, managing regional networks and being strongly connected to research.
Furthermore, the combination of the above-mentioned activities, together with regional
participation in national planning and strategy for chlamydia prevention, could be a vital
part of the prevention mix.
QUALITY ENHANCEMENTS AND QUALITY ASSURANCE OF LABORATORY DIAGNOSIS
OF SEXUALLY TRANSMITTED INFECTIONS IN EASTERN EUROPE
M. Domeika,1 R Ballard,2 and M Unemo3 on behalf of the Eastern European Network for
Sexual and Reproductive Health (EE SRH Network)
Department of Medical Sciences, Uppsala University, Uppsala, Sweden
(marius.domeika@medsci.uu.se)
2
Division of STD Prevention, Centers for Disease Control and Prevention (CDC), Atlanta, USA
3
Swedish Reference Laboratory for Pathogenic Neisseria, Department of Laboratory
Medicine, Clinical Microbiology, Örebro University Hospital, Örebro, Sweden
1
Background. Sexually transmitted infections (STIs), excluding HIV, are not considered as
major burden of public health in Eastern European countries and therefore are left aside,
e.g. mainly no national strategies or programs exist, and laboratory diagnosis, patient
management and epidemiological surveillance are suboptimal.
Initial goal: To optimize, harmonize and quality assure the laboratory diagnosis of STIs in the
catchment countries.
Methods. Through participation in the multinational and multidisciplinary Eastern European
Network for Sexual and Reproductive Health (EE SRH Network), presently including active
representatives from 14 EE countries as well as international STI experts from EU countries
and USA, to reach consensus on ideal performance and logistics for the laboratory diagnosis
of STIs with further implementation and legalization of the materials elaborated on country
level.
ABSTRACTS
Results. Following consensus has been reached: sole use of microscopy for the diagnosis of
N gonorrhoeae should be limited to the diagnosis of male symptomatic urethritis and use of
culture and NAATs have to be promoted; surveillance of antimicrobial resistance is crucial to
establish. NAATs have to become the method of choice for the diagnosis of C trachomatis
and M genitalium, and wider implemented for diagnosis of T vaginalis, syphilis and herpes
infections. Serology for diagnosis of C trachomatis and T vaginalis infections has to be
abolished. Wet smear microscopy is still recognized as a sufficient tool for the diagnosis
of T vaginalis and bacterial vaginosis (BV). BV can also be diagnosed using a Gram-stained
smear. NAATs that are not internationally acknowledged should be validated against the
international standard assays. International and nationally adjusted guidelines have been
produced and published (English and respective national languages).
Conclusion. Adoption of internationally acknowledged, evidence-based standards will allow
countries improvement of their medical care standards. Since its formation, the EE SRH
Network has been effective in facilitating this process.
22
DETECTION OF CHLAMYDIA TRACHOMATIS BY MOLECULAR BIOLOGICAL
METHODS IN LATVIA
Corresponding author: Diana Dusacka
Other authors: Aija Gulbe, Natalia Repuscenko, Natalia Novikova, Violeta Mavcutko, Tatjana
Kolupajeva, Jelena Storozenko
Institution of corresponding author: State Agency "Infectology Center of Latvia"( ICL)
Institutions of other authors: State Agency "Infectology Center of Latvia"( ICL)
Speciality: microbiologist
Country: Latvia
E-mail: diana.dusacka@lic.gov.lv
Introduction: Chlamydia trachomatis is a mandatory notifiable infection in Latvia. Different
diagnostic methods for detection of C. trachomatis were used in Latvia. In early 1990s
Chlamydia serology and Chlamydia antigen detection assays - DFA (Direct Fluorescent
antibody test) and EIA (Enzyme Immune Assay) were widely used. Since 1996s some
laboratories started Chlamydia detection by DNA probe assay and Nucleic Acid Amplification
Technique (NAAT).
Objectives: The aim of this study is to give short overview of methods, currently used in
Latvia and to analyze experience of testing by molecular biological technologies.
Methods: Data from the national computer-based surveillance system VISUMS and ICL
laboratory data were used to analyze laboratory methods for confirmation of Chlamydia
infection in Latvia.
Discussion: According to published data, NAAT are the most sensitive (90-95%) tests for
genital Chlamydia (DFA sensitivity: 65-75%). In spite of this fact, in Latvia NAAT is not widely
used. Possible reasons are: expensive equipment and reagents, special rooms requirements,
need for high personal skills. New technology validation has to be continued, however
preliminary ICL data showed approximately the same rate of positive results by DFA and
NAAT in a limited group of samples. DFA results can be very sensitive due to experience of
microscopist, but the method is not suitable for large workloads.
Conclusions: 474/1042 (45.5%) cases of Chlamydia infection registered in 2010 were
confirmed by molecular biological methods, but only 9.4% - by NAAT. High sensitivity and
specificity of NAAT tests, simultaneous detection of several agents (e.g. C. trachomatis and
N. gonorrhoeae), possibility to use non invasive clinical samples for testing, make it suitable
for improvement of C.trachomatis diagnosis, and for large volume screening.
23
ABSTRACTS
Results: 1042 laboratory confirmed cases of Chlamydia infection were registered in Latvia in
2010: 329 (31.6%) positive results were identified using DFA, 378 (36.3%) - DNA probe assay,
96 (9.4%) - NAAT. C.trachomatis DFA method traditionally was mostly used in ICL laboratory.
In 2010, 3829 and 126 patients were tested by DFA and by DNA Probe assay, respectively.
Frequency of positive results for C. trachomatis was 5% and 6.3%, respectively. A NAAT based
method (Aptima Combo 2 Assay Gene-Probe), was recently introduced in ICL. Parallel testing
of clinical specimens (swabs, urine) from 100 patients were performed by both methods:
6/100 were positive by DFA and 7/100 by NAAT.
PREVALENCE OF HIGH-RISK HUMAN PAPILLOMAVIRUS INFECTION IN CERVIX
CYTOLOGIC SMEARS IN ALBANIA
Corresponding author: Kozeta Filipi1
Other authors: Federico De Marco2, Gentian Kusta3, Mirela Rista4, Albana Ahmeti1.
Institution of corresponding author:
1. Institute of Public Health, Department of Epidemiology, Tirana, Albania
Institutions of other authors:
2. Laboratory of Virology – “Regina Elena” Cancer Institute, Rome, Italy
3. Maternity Hospital, Berat, Albania
4. Maternity Hospital, Tirana, Albania
Speciality: epidemiologist
Country: Albania
E-mail: kkozeta@hotmail.com
Introduction: Cervical cancer ranks the 2nd most frequent cancer in women in Albania,
and the 2nd most frequent cancer among women between 15 and 44 years of age (IARC,
GLOBOCAN 2008). The established cause is high-risk types of human papillomavirus (HPV),
and a new modality for cervical cancer screening is the combination of cervical cytology with
HPV testing.
Objectives: To identify the prevalence of HPV infection, the types of HPV and cervical
cytological profiles of healthy Albanian women.
Methods: This cross-sectional investigation of HPV typing together with cytology screening
in Albania was conducted between 2004 and March 2007, covering a total of 402 women.
HPV detection and typing were carried out by a combined MY09/MY11 and GP5+/GP6+ PCR
followed by direct sequencing of generated amplicons.
ABSTRACTS
Results: The mean age of the study group was 36 years (range 19-75 years) and the
prevalence of high risk HPV infection was 15%. This figure was higher (25.2%) among women
aged between 19-29 and lower (13.6%) among those aged 30 or over. HPV 16 was found
to be the most frequent type (41% of cases), followed by HPV 53 (7.2%), HPV 31 (5.8%), and
HPV 18 (4.3%). HPV 81 and HPV 84 were the most prevalent low-risk types detected with
prevalence of 11.6% and 5.8%, respectively.
Discussion: The use of a simple HPV DNA test followed by immediate ‘screen and treat’
algorithms based on visual inspection in those who are HPV positive are needed to minimize
the number of visits and make best use of limited resources. Detailed knowledge of HPV
type circulating patterns in specific local geographical areas is essential for appropriate
implementation of screening, prevention, and surveillance campaigns.
Conclusions: In this study no differences were noted in types of HPV between young and
mature women. The circulation of HPV types is far more complex than assumed generally.
24
CHALLENGES AND FACTS ON HPV VACCINATION
IN ROMANIA
Corresponding author: Viorica Gheorghiu 1*
Other authors: Aurora Stanescu, Alina Zaharia, Denisa Janta, Teodora Solomon 1*
Institution of corresponding author: National Center for Surveillance and Control of
Communicable Diseases 1*
Institutions of other authors: National Center for Surveillance and Control of Communicable
Diseases 1*
Speciality: public health and epidemiology
Country: Romania
E-mail: viorica.gheorghiu@insp.gov.ro
Introduction: Despite no specific serological survey regarding the prevalence of the HPV
carriers being conducted in the last 10 years, the decision makers and those responsible from
the Ministry of Health have decided to introduce the HPV vaccination in November 2008. The
target group was girls aged12-13 years. The implementation of this vaccination campaign
had many constraints.
Objectives: Identification of knowledge, attitude and behavior which led to acceptance or
rejection of the 2 types of HPV vaccines (Cervarix and Silgard) by children from the target
group or/and their tutors.
Methods: The National Center for Information and Prevention of Cervix Cancer organized
with the help of National Center for Surveillance and Control of Communicable Diseases an
information campaign between 15.05.2009 - 18.12.2009 using a "green line" phone and an
electronic information site.
Discussion: After the implementation of information campaign the acceptability of the
HPV vaccine improved but still there are many who refused the second or third doses.
For avoiding the wastage of the vaccine the population target group was extended up to
24 years of age. The HPV vaccination campaign had a negative impact on routine
vaccination too.
Conclusions: The introduction of the new vaccines is more and more difficult to be accepted
by general population and the information campaign for HPV vaccine should to precede and
start at least 4 months before vaccination itself.
25
ABSTRACTS
Results: During the six months of the campaign a total of 3626 phone calls and 362 emails
were recorded. From this huge number, only 1955 calls were specific for cervical cancer and
HPV infection. All the questions have been coded and then analyzed. The main topics of the
questions were focused on: information regarding the development and implementation
of the campaign (17,57%), information about the vaccines used (33,79%), information
regarding cervical cancer (48,69%).
EPIDEMIOLOGY OF CHLAMYDIA TRACHOMATIS INFECTIONS IN GERMANY:
DATA FROM STD-SENTINEL
Corresponding author: Karin Haar
Haar K (1), Bremer V (2), Sailer A (1), Marcus U (1), Hamouda O (1)
(1) Department for Infectious Disease Epidemiology, HIV/AIDS, STI and Bloodborne
Infections Unit, Robert Koch-Institute, Berlin, Germany
(2) EPIET Chief coordinator, European Centre for Disease Prevention and Control (ECDC),
Stockholm, Sweden
Country: Germany
E-mail: HaarK@rki.de
Introduction: Chlamydia trachomatis (CT) infections are often asymptomatic, especially
in women, and can lead to infertility. In Germany, the new Infection Protection Act was
implemented in 2001 and only syphilis, HIV and hepatitis were stated reportable STIs.
A STD sentinel surveillance system was set up by the end of 2002, where a subset of health
care providers voluntarily reported the incidence of STIs. Further, in 2008 an opportunistic
chlamydia screening programme for all women under the age of 25 was introduced in
Germany.
Objectives: To collect data on the frequency of STIs, the demographic features and
geographic distribution, a clinical STD-sentinel surveillance system was implemented in a
country where most STIs are not reportable. We tried to identify risk groups and behaviour to
improve guidance on targeted prevention and future surveillance initiatives.
Methods: Between the end of 2002 and 2009 local health authorities, specialized outpatient
clinics and practitioners continually reported examination and infection data, demographic
characteristics and possible risk factors for STI-acquisition. Patients were asked to return
completed questionnaires which were linked to doctors’ reports and provided further
information about patients’ social state, sexual behaviour and possible source of infection.
ABSTRACTS
Results: Of all chlamydia-examinations, 6% (5955/98405) were positive. This was higher than
for any other lab-confirmed STI, namely gonorrhoea (GO) 3.6%, HIV 1.1%, syphilis 3% and
trichomonas 2.5%.
For 2640 chlamydia cases (64% women) further data were provided: The overall mean age
was 26 years, men being significantly older. 63% of women were migrants, 70% sex workers
and 45% of men were MSM. 13% of women had CT in the past, 17% of men had a previous
episode of GO. In total, 10% were co-infected with GO.
54% of women stated “health check” as their reason of attendance, whereas 23% reported
“health problems”. 35% of women believed their regular partner to be the source of infection,
33% named customers, 61% of men reported casual partners.
The median number of sexual partners within the last 6 months was 3 in men and 2 in
women. 23% of men and 47% of women reported consistent, 32% of men and 6% of women
reported no condom use with casual partners.
26
Discussion: Due to high rates of CT-infected women who only attended for a “health check”
i.e. were asymptomatic, CT might be an underdiagnosed but frequent infection in Germany.
For countries with lacking STI-notification systems, implementation of an STD-sentinel seems
feasible; however, data have to be interpreted cautiously as selection bias is always an issue.
Patients seen in STD-Sentinel institutions have high re- and co-infection rates, are frequently
working in the sex industry and are migrants or MSM. Therefore data from the sentinel
cannot be interpreted as infection rates in the general population.
Conclusion: Clinical STD-sentinel surveillance systems are useful to match laboratory
findings with behavioural data; however, recruitment sites have to be chosen according to
the studied core group. The German CT screening programme is now accompanied by
CT-laboratory sentinel surveillance. Data from this lab sentinel will provide more reliable
public health data on the epidemiology of chlamydia in Germany.
EPIDEMIOLOGICAL KNOWLEDGE BY GENOTYPING CHLAMYDIA TRACHOMATIS:
AN OVERVIEW OF RECENT ACHIEVEMENTS
Corresponding author: Björn Herrmann
Institution of corresponding author: Department of clinical microbiology, Uppsala University
Hospital, Sweden
Country: Sweden
Email: bjorn.herrmann@medsci.uu.se
Overview: Several C. trachomatis typing methods based on other target genes than ompA
have been published in recent years. Two multilocus sequence typing (MLST) schemes based
on housekeeping genes have been shown useful for analysis of evolutionary changes and
slowly evolving processes [1, 2]. Their differentiating capacity is similar to ompA sequence
analysis and therefore they have limitations in applications for short term epidemiogy.
A third MLST system was developed by Klint et al. [3] and is intended for clinical
epidemiology and outbreak investigations. It is based on five highly variable but stable
genomic loci which give the system up to a fivefold higher resolution than ompA. A variety
of clinical specimens have been analysed, including urogenital chlamydia [4-6], LGV [7] and
trachoma [8]. There is also a multilocus variable number tandem repeat (VNTR) analysis
(MLVA) system which offers a resolution similar to that of the MLST system by Klint et al.
[9, 10]. However, both MLST and MLVA require several days of work until final results are
achieved and they are expensive. Diagnostic DNA microarray technology has emerged as an
alternative in microbial genotyping and reduces the cost and the work load. An ompA array
has been published [11], but still no high resolution array for C. trachomatis.
27
ABSTRACTS
Background: Chlamydia trachomatis is one of the most common sexually transmitted
infections world-wide. Typing of C. trachomatis is important to understand its epidemiology
and thereby decrease the prevalence. Conventional genotyping has been predominated by
analysis of the major outer membrane protein or the encoding ompA gene. However, the
differentiating capacity of ompA analysis is limited and methods with higher resolution are
needed to perform adequate short term epidemiological analyses.
Summary: Genotyping methods for C. trachomatis have been improved in recent years and
conventional ompA sequencing has limited use compared to current alternatives.
References:
1. Pannekoek et al. Multi locus sequence typing of Chlamydiales: clonal groupings within
the obligate intracellular bacteria Chlamydia trachomatis. BMC Microbiology 2008;8:42.
2. Dean et al. Predicting phenotype and emerging strains among Chlamydia trachomatis
infections. Emerg Infect Dis. 2009;15:1385-94.
3. Klint et al. High-resolution genotyping of Chlamydia trachomatis strains by multilocus
sequence analysis. J Clin Microbiol 2007;45:1410-4.
4. Herrmann et al. Emergence and spread of Chlamydia trachomatis variant, Sweden.
Emerg Infect Dis 2008;14:1462-5.
5. Jurstrand et al. Characterisation of Chlamydia trachomatis by ompA sequencing and
multilocus sequence typing in a Swedish county before and after identification of the
new variant. Sex Transm Infect 2010;86:56-60.
6. Christerson et al. Multilocus Sequence Typing of Urogenital Chlamydia trachomatis From
Patients With Different Degrees of Clinical Symptoms. Sex Transm Dis 2011; epub Feb 4.
7. Christerson et al. Typing of lymphogranuloma venereum Chlamydia trachomatis strains.
Emerg Infect Dis 2010;16:1777-9.
8. Harding-Esch et al. Multi-locus sequence typing: a useful tool for trachoma molecular
epidemiology. In: Stary et al (eds). Chlamydial infections Proceedings of the 12th
International Symposium on Human Chlamydial Infections; Salzburg 2010. p. 55-8.
9. Pedersen et al. Highly discriminative genotyping of Chlamydia trachomatis using omp1
and a set of variable number tandem repeats. Clin Microbiol Infect 2008; 14:644-52.
10. Wang et al. Evaluation of a High Resolution Genotyping Method for Chlamydia
trachomatis Using Routine Clinical Samples. PloS one 2011, 6(2):e16971.
11. Ruettger et al. Genotyping of Chlamydia trachomatis strains from culture and clinical
samples using an ompA-based DNA microarray assay. Molecular and cellular probes
2010;25:19-27.
LABORATORY SURVEILLANCE OF CHLAMYDIA TRACHOMATIS
IN DENMARK 1994-2010
Corresponding author: Steen Hoffmann
Institution of corresponding author: Statens Serum Institut, Artillerivej 5 DK-2300
Copenhagen S
ABSTRACTS
Speciality: Clinical microbiology
Country: Denmark
E-mail: hof@ssi.dk
Introduction: The Danish laboratory surveillance system for oculo-genital infections caused
by Chlamydia trachomatis (CT) was introduced in 1988 and became mandatory in 1994. This
presentation will give an overview of the design of the surveillance system as well as some
of the results, and future perspectives and challenges.
28
Objectives: The objective of the surveillance system is to monitor the occurrence of
laboratory diagnosed chlamydia in Denmark.
Methods: Diagnosis of CT in Denmark is performed at 15 public laboratories, 13 in clinical
microbiology and two in clinical biochemistry. They use five different laboratory data
systems. The laboratories report quarterly the number of tests performed and the number of
diagnosed patient cases. For each case, the following variables are reported (anonymously):
age, gender, specimen date, anatomical site, type of health care provider, and laboratory
method used. There is no clinical reporting.
Results: From 1994 to 2009 the annual number of analyses increased from approximately
270,000 to nearly 350,000 and the number of laboratory diagnosed cases of chlamydia
increased from 14,000 to 30,000. In 2009, the incidence among males was 414 per 100,000
and it was 665 among females; 83% and 90%, respectively, were 15-29 years old. The
incidence among females aged 18 years appears to be increasing. The fraction of males
among all diagnosed cases has slowly increased from 23% to 38%. Almost 95% of all
specimens were obtained by general practitioners. Since 2003 NAATs were used in more than
99% of cases. Urine is increasingly being used as specimen material, especially among males.
Discussion: The strength of this surveillance system is that it is mandatory and nationwide.
The weakness is that no data are reported regarding persons with negative tests. Thus, it is
not possible to account for the examined population, e.g. in terms of age, gender, anatomy
or indication for testing. However, a nationwide Microbiology Data Bank is currently being
developed to receive real-time complete data for all positive and negative microbiology
laboratory test results, including chlamydia tests.
Conclusions: This surveillance system provides reliable information about the number of
laboratory confirmed cases of chlamydia. As of 2007, the responsibility for interventions
to reduce chlamydia was transferred from the counties (n=15) to the municipalities
(n=101). This has increased the need for more detailed information about the geographical
distribution of chlamydia. Also, data about the age distribution of those examined must be
established in order to allocate resources more cost-effectively.
SURVEILLANCE OF CHLAMYDIA TRACHOMATIS IN ENGLAND:
A REGIONAL PERSPECTIVE
Corresponding author: Charlie Jennison
Other authors: Lynsey Emmett Iain Roddick Adrian Deas Sam Bracebridge
ABSTRACTS
Institution of corresponding author: Health Protection Agency
Institutions of other authors: Health Protection Agency
Speciality: Chlamydia surveillance
Country: England
E-mail: charles.jennison@hpa.org.uk
29
Introduction: In England genital C. trachomatis testing can be broken into three main
categories. Testing that occurs in: Genitourinary Medicine (GUM) clinics, the National
Chlamydia Screening Programme (NCSP) and "Community testing" outside these two
specific services. GUM and NCSP data are collected as well defined disaggregate datasets,
with community testing as aggregate data by most regions (6/9). In the East of England,
disaggregate community testing data have been collected centrally since 2008.
Objectives: To collect a disaggregate community testing dataset for the East of England and
consequently combine this with the other available datasets for the East of England, covering
a time period of 30 months for the 15-24 year age group. This would allow for further
investigation into chlamydia testing in the region. The combined dataset would be queried
to assess progress towards achieving testing levels needed for control.
Methods: Data extracts from the Genitourinary Medicine Clinic Activity Dataset (GUMCAD),
the NCSP database and the East of England community testing database were combined in
SQL Server 2005 and then analysed using STATA (version 11).
Results: Results from Jennison et al. (Journal of Public Health, 2011), will be presented along
with results from the further developmental analyses.
Discussion: Combining and standardising the three datasets was possible and has resulted
in the most complete surveillance data of C. trachomatis testing in England currently
available. There are several benefits of this dataset, for example it allows for the easy
answering of commisioners' questions or the breakdown of testing by; date of attendance,
geographic location, age and sex. The benefits and limitations will be expanded upon in
this presentation.
Conclusions: Recent improvements in data capture for chlamydia testing in England have
resulted in excellent data sources for further epidemiological analyses. In combining two
of these datasets enhanced surveillance reports for the East of England have already been
used to inform service provision and help commissioners meet Chlamydia testing coverage
targets. The dataset produced in this work takes this a step further and can be used to look
in depth at a more complete picture of chlamydia epidemiology.
PREVALENCE AND GENETIC STRUCTURE OF CHLAMYDIA TRACHOMATIS
IN ESTONIA
Corresponding author: Kai Joers
Institution of corresponding author: Quattromed
ABSTRACTS
Country: Estonia
E-mail: Kai.Joers@Laborid.ee
Chlamydia trachomatis is one of the most common of pathogens causing sexually
transmitted diseases. In the lecture I describe the prevalence of C. trachomatis in Estonia and
its population structure based on samples originating from gynecologists and andrologists.
The general prevalence of C. trachomatis among Estonian population is 7,3 %, based on
analysis of 1200 samples.
30
The most abundant C. trachomatis serotypes in Estonia were serotype E (32,6 %), F (17,4 %),
K (15,2 %), G (13,0 %), D (8,7 %), J (6,5), B (4,3 %) and H (2,2 %). This is very similar to serotype
prevalence in Sweden. As one third of all samples belonged to serotype E, serotyping only
has not enough discriminatory power to separate possibly relevant different strains.
For better characterization of Estonian C. trachomatis population genotyping method based
on Multi Locus Sequence Typing (MLST) was introduced. It was found that MLST based on
8 loci had a discriminatory index D = 0.97 and thus suitable for genotyping for clinical use.
15 unique genotypes were identified among 21 clinical samples, indicating the existence of
heterogeneous population with probably long history.
PREDICTION ON TUBAL FACTOR INFERTILITY BY MEASURING C. TRACHOMATIS AND
CHLAMYDIAL HEAT SHOCK PROTEIN 60-SPECIFIC ANTIBODY AND CELL-MEDIATED
RESPONSES – A PROSPECTIVE STUDY
Corresponding author: Päivi Joki-Korpela
Other authors: Tiina Rantsi, MD Christel Hyden-Granskog, PhD Jorma Paavonen, MD, PhD,
Prof Tiina Rantsi, MD Heljä-Marja Surcel, PhD Aila Tiitinen, MD, PhD, Prof Hanna Öhman, MSc
Institution of corresponding author: Department of Obstetrics and Gynecology, Helsinki
University Central Hospital
Institutions of other authors: Department of Obstetrics and Gynecology, Helsinki University
Central Hospital Institute for National Health and Welfare, Oulu
Speciality: Gynecology and obstetrics
Country: Finland
E-mail: paivi.joki-korpela@helsinki.fi
Introduction: Infertility is a common problem, and examinations and treatments for patients
are costly. Approximately one third of infertile couples suffer from tubal factor infertility (TFI).
Non-invasive methods for detecting TFI are needed for effective and economical planning
of the treatment. Our group has shown retrospectively that TFI prediction model could be
improved by combining four distinct markers for humoral and cell-mediated immunity.
Methods: C.trachomatis and CHSP60-specific immunoglobulin G plasma antibodies have
been analyzed by ELISA. Antibodies against CHSP-60 were measured using recombinant
CHSP60 as an antigen. Cell mediated immunity has been studied by using peripheral blood
mononuclear cells. C.trachomatis EB; serovars E and F, and recombinant CHSP 60 have been
used as antigens.
Results: So far, we have collected and analyzed blood samples from approximately 225
couples (450 patients), and it seems that approximately 9% of patients (4.5% couples, 2.4%
man only, 1.8% woman only) are positive with these markers. We are collecting data on
31
ABSTRACTS
Objectives: The aim is to validate the developed TFI prediction model in order to develop
sensitive non-invasive test to be able to efficiently and economically plan further treatment
for subfertile couples.
patients (results on examinations and treatments, age, previous pregnancies, STDs, illnesses,
medication, BMI, smoking, use of alcohol) and will analyze and compare the obtained data
with results on chlamydial immune response.
Conclusions: Subfertility can be caused by several factors e.g. male factors, endometriosis,
hormonal factors, TFI and unexplained infertility. Laparoscopy is a golden standard when
detecting TFI. However, it is invasive, costly and can cause complications to patients.
Moreover, it's unable to tell the essential function of the ovarian tubes. Non-invasive
methods are needed to identify TFI for effective and economical planning of the treatment.
DETECTION OF CHLAMYDIA TRACHOMATIS BY GENETIC METHODS IN A SWEDISH
COUNTY BEFORE AND AFTER IDENTIFICATION OF THE NEW MUTATED VARIANT
Corresponding author: Margaretha Jurstrand
Other authors: Björn Herrmann (BH), Hans Fredlund (HF), Magnus Unemo (MU)
Institution of corresponding author: Clinical Research Centre, Örebro University Hospital,
SE-70185 Örebro, Sweden
Institutions of other authors: Department of Clinical Microbiology, University Hospital,
SE-751 85 Uppsala, Sweden.(BH) Department of Laboratory Medicine, Clinical Microbiology,
Örebro University Hospital, SE-70185 Örebro, Sweden. (HF, MU)
Speciality: PhD, Laboratory Manager
Country: Sweden
E-mail: margareta.jurstrand@orebroll.s
Introduction: Despite mandatory partner notification in Sweden legislated in 1988, the
incidence of Chlamydia trachomatis has increased each year since 1997. In 2006 a new
variant of Chlamydia trachomatis (nvCT), with a deletion in the cryptic plasmid, was reported
in Sweden. This deletion included the targets for the genetic diagnostic systems used in
many clinical laboratories and resulted in thousands of false negative results.
ABSTRACTS
Objectives: To study the distribution of C. trachomatis omp1 genotypes within sexual
networks and to determine if genotyping would improve partner notification. The aim was
also to characterize genetic variation among wild genotype E isolates and among nvCT
isolates using a high-resolution multi locus sequence typing (MLST) system.
Methods: A PCR method was developed for characterization of clinical C. trachomatis
isolates by sequence analysis of the omp1 gene. The method was used for characterization
of genotypes in consecutive Chlamydia tissue culture positive samples from late 2006, and
was compared with the results of a one-year study performed from 1999 to 2000 in the same
area. Further on we used a high-resolution multi locus sequence typing (MLST) system for
analysis of C. trachomatis genotype E isolates found in 2006.
Results: Sequence analysis of the omp1 gene revealed that the most prevalent genotypes
was C. trachomatis genotype E, followed by F, D, K, Ba, G, H, I, and J in 1999-2000. In the
largest network comprising 26 individuals three different C. trachomatis genotypes were
found. The distribution of genotypes was similar in year 2006 but a significant increase
32
of genotype E from 47% in year 1999-2000 to 69% in 2006 was detected. Complete MLST
sequence types (STs) were obtained from 24 of 28 wild type C. trachomatis genotype E
isolates and among these 14 (58%) unique STs were identified. In these 24 isolates four omp1
variants were detected. In contrast all nvCT isolates (n=41) displayed an identical ST.
Conclusions: Genotyping was shown to be useful in a partner notification context, but
was found to provide limited discrimination especially for the predominating genotype E.
Using the MLST system the discrimination of C. trachomatis genotype E wild type isolates
is considerable higher compared to omp1 sequencing, which enables improved studies
in molecular epidemiology. All nvCT in 2006 was of identical genotype E which strongly
indicate a clonal spread of the nvCT.
EVALUATION OF CHLAMYDIA SCREENING IN DIFFERENT COMMUNITY SCREENING
VENUES IN LAMBETH, LONDON, UK 2005-2010
Corresponding author: Dr Sebastian Kalwij
Other authors: Rumbi Mugezi
Institution of corresponding author: Amersham Vale Practice, London, UK
Institutions of other authors: Lambeth and Southwark Chlamydia Screening Office
Speciality: Primary Care
Country: England
E-mail: sebastiankalwij@mac.com
Introduction: Lambeth Primary Care Trust in London was one of the first PCT’s in England,
to offer screening to young patients (age 16-25), in a variety of community settings. The
programme is now in its seventh year. It is well established and screens large numbers
of young people each year. We tried to map the screening activity and identify screening
venues who screen the most and reach those most at risk.
Methods: Most screening venues used the two main laboratories at Kings College Hospital
and St Thomas Hospital. Tests requested through the website (www.checkurself.org.uk) were
sent to a private laboratory (TDL, The Doctors Laboratory) in Central London. Each venue has
a so-called site-code and were grouped as follows: general practice, community reproductive
and sexual health clinics, Brook Advisory (Charity aimed at young people, contraception and
STI’s) pharmacies, midwives, outreach services and the website. These figures were collected
on a quarterly basis every year since 2005. This study excluded invalid screens and all tests
from Genito-Urinary Hospital Clinics as stipulated by the National Chlamydia Screening
Programme.
Results: 33, 317 screens where processed over 5 years (1/4/2005-31/3/2010) at the three
laboratories and the results collected by the local Chlamydia Screening Office. The results
were broken down by screening venues; General practice 15928 (47.8%), Community
33
ABSTRACTS
Objectives: The objective is to find out which community screening venue contributes the
most in both volume and positivity rate. This is an important question to answer in times of
economic uncertainty and health care budget cuts.
Reproductive and Sexual Health Clinics 7235 (21.7%), Brook Advisory 6911 (20.7%),
Pharmacies 773 (2.3%) and midwives, outreach programmes and the website contributed
2470 (7.4%). During the same period the number of tests for chlamydia across all venues
increased from 3745 tests in 2005/06 to 10.390 tests in 2009/10. Positivity rates were as
follows: General Practice 7.1%, Community Reproductive and Sexual Health Clinics 11.7%,
Brook Advisory 5.3%, Pharmacies 6.7%, Midwives 6.3%, Website: 4.8%. Overall positivity rate
for 2009/2010 was 7.9%.
Discussion: Whilst chlamydia prevalence remains high (6-9% in certain inner city areas) it is
important to identify a practical approach to screen large number of asymptomatic young
people year after year. Whilst certain technological innovations like internet testing sound
promising, it may not replace the traditional face-to-face consultation with a trusted Health
Care Professional in either Community Reproductive and Sexual Health Clinics or General
Practice yet.
Conclusions: In Lambeth PCT, General Practices and the Community Reproductive and
Sexual Health Clinics were responsible for the vast majority of Chlamydia tests. This
demonstrates that both service providers are vital in offering the Chlamydia screening
service. To set up and sustain a screening programme it is important to offer opportunistic
screening via venues such as general practices and Community Reproductive and Sexual
Health Clinics as they see large number of young people coming through their doors and
detect those most at risk.
8 YEARS OF CHLAMYDIA SCREENING IN ENGLAND; EXPERIENCES AND
LESSONS LEARNT
Corresponding author: Dr Sebastian Kalwij
Institution of corresponding author: Amersham Vale Practice, New Cross, London.
Speciality: Primary Care
Country: United Kingdom
E-mail: sebastiankalwij@mac.com
Introduction: This presentation aims to give an overview of the National Chlamydia
Screening Programme in England, which is now in its 8th year. The programme has been
set up as an opportunistic screening programme and chlamydia screening is offered to all
sexually active young people between the ages of 15 and 24 years in all 152 Primary Care
Trusts in England.
ABSTRACTS
Objectives: To identify a successful strategy to screen 35% of 15-24 year olds in England.
Methods: I looked at data collected by the Health Protection Agency from 2003 to 2011.
Results from all laboratories participating in the programme and the Chlamydia Screening
Offices are collected by the Health Protection Agency on a quarterly basis. These figures get
published in the annual reports.
Results: The number of tests has increased from 18.529 in 2003 to 1.2 million in 2010/11
(22.1% coverage). The most successful screening venues in terms of volume (n=975.989)
are: Outreach: 23.1%, Community Sexual Health Clinics: 21.7%, General Practice: 16.5%.
34
Alternative settings like internet testing: 2.7% and testing in local pharmacies 2.2%, reached
far fewer people. In terms of positivity rate, the picture looks different. Community Sexual
Health Clinics: 8.0%, Internet testing 7.8%, Pharmacy: 7.1%, General Practice: 5.5% and
Outreach: 2.9%.
Discussion: In order to roll out a Chlamydia Screening Programme it is important to identify a
successful strategy to reach high risk populations. Whilst established clinics and primary care
services are already seeing many young people, novel approaches are needed to reach those
who don't visit these health care facilities on a regular basis.
Conclusions: It is feasible to screen a large percentage of the target population per year. A
diverse approach is needed to reach those at high risk for chlamydia whilst covering a high
percentage of the target population in order to make an impact on controlling Chlamydia
infection and reducing its prevalence.
DEVELOPMENT OF AN OMPA PYROSEQUENCING ASSAY FOR CHLAMYDIA
TRACHOMATIS BASIC GENOTYPING DIRECTLY FROM UROGENITAL AND
OCULAR SPECIMENS
Corresponding author: Rok Kogoj
Other authors: Potocnik Marko, Darja Kese
Institution of corresponding author: University of Ljubljana, Medical Faculty, Institute of
Microbiology and Immunology, Zaloska 4, 1000 Ljubljana, Slovenia
Institutions of other authors: Potocnik M.: University Medical Centre Ljubljana, department
of Dermatovenereology, Zaloska 2, 1000 Ljubljana, Slovenia; Kese D.: University of Ljubljana,
Medical Faculty, Institute of Microbiology and Immunology, Zaloska 4, 1000 Ljubljana,
Slovenia
Speciality: Intracellular bacteria
Country: Slovenia
E-mail: rok.kogoj@mf.uni-lj.si
Objectives: The aims of this study were to develop and validate a procedure for the genetic
characterization of clinical C. trachomatis strains by a newly developed pyrosequencing
assay that targets the variable domains VD I and VD IV of the ompA gene and to define the
prevalence of particular genotypes among symptomatic Slovenian sexually active adults.
Methods: A combination of three pyrosequencing assays that cover the whole VD-I and
a part of the VD-IV was optimised with eleven reference C. trachomatis strains (D-L3) and
validated by comparison to ompA Sanger sequencing. Additionally, 74 urethral swabs from
men, 78 cervical swabs from women and 31 ocular swabs, which were previously diagnosed
as C. trachomatis positive, were evaluated by pyrosequencing, with their results compared
to the results obtained by Sanger sequencing.
35
ABSTRACTS
Introduction: In epidemiological studies, Chlamydia trachomatis is mainly genotyped
by conventional sequencing of the ompA gene which requires long turnaround times,
lots of hands on time and is relatively high cost. To our knowledge, pyrosequencing, a
bioluminimetric »de novo« sequencing technique, has not yet been tested as a tool for
C. trachomatis genotyping.
Results: OmpA VD-I forward, VD-I reverse and VD-IV pyrosequencing products were obtained
from all reference C. trachomatis strains and all patients' samples that were positive in the
respective pyrosequencing PCR. After software analysis VD-I forward, VD-I reverse and
VD-IV pyrosequencing products showed average "good quality" lengths of 74, 74 and
80 nucleotides respectively and a 100% match to Sanger sequences. The most prevalent
genotype was E (53.4%) followed by F (19.9%), D (7.5%), K (6.8%), G (6.2%), H (3.1%), J (1.9%)
and Ia & Ja (0.6%).
Discussion: Pyrosequencing the ompA gene could be used as a tool for genotyping
C. trachomatis in epidemiological purposes. The main concern still remains its inability to
produce more than maximally 100bp sequence reads. However, when sequencing carefully
selected key areas of a gene, it is possible to achieve satisfactory amounts of sequence data
to differentiate between genotypes. In our study population genotype E was shown to be
dominant which is in concordance with several studies all over the world.
Conclusions: Pyrosequencing is a rapid and easy to perform method; it is economically
effective and could be used as an alternative to conventional sequencing for C. trachomatis
epidemiological purposes. Our study contributes to the creation of an informative
C. trachomatis genotyping protocol, which could enable rapid strain identification and
facilitate C. trachomatis genotypes distribution studies in different patients groups.
POOLING GENITAL SAMPLES COULD BE A COST BENEFICIAL APPROACH FOR
SCREENING CHLAMYDIA TRACHOMATIS IN YOUNG FEMALE PATIENTS IN LITHUANIA
Corresponding author: Vesta Kucinskiene1
Other authors: Skaidra Valiukeviciene1, Marius Domeika2
Institution of the corresponding author: 1Department of Skin and Venereal Diseases, Medical
Academy, Lithuanian University of Health Sciences, Kaunas, Lithuania.
Institution of other authors: 2Department of Medical Sciences, Uppsala University, Uppsala/
Eastern European Committee of Swedish Health Care Community, Stockholm, Sweden.
Speciality: Dermatovenereologist
Country: Lithuania
ABSTRACTS
E-mail: kvesta@delfi.lt
Introduction: Laboratory diagnostic tests to detect C. trachomatis are expensive and not
accessible for every young person in Lithuania. That is one of the reasons why the sample
size of testing for C. trachomatis is very low in Lithuania though epidemiological studies
show the infection to be prevalent, eg. the prevalence among 15-19-year old sexually active
lithuanian adolescent girls is 18.2%.
Objectives: to estimate the prevalence of Chlamydia trachomatis infection among young
women (20-24 year old); to estimate the accuracy of pooling vaginal samples; to compare the
costs of DNA hybridization method to detect C. trachomatis, when analyzing individual and
pooled vaginal samples.
36
Methods: A total of 795 female students aged 20-24 from Kaunas high schools were invited to
participate in the study. The response rate was 67 %. Study participants were surveyed using a
standardized questionnaire, and collected the vaginal swabs themselves following the written
instructions. All vaginal samples were prepared and analyzed using Digene Hybrid Capture II
CT/NG Test in accordance with the instructions of the manufacturer. Specimens were tested
individually and in thoroughly mixed pools comprising 3 consecutive samples. For validation
of positive results, all positive individual samples and pools were retested. According to the
test results, the sensitivity, specificity and cost savings of pooling samples were assessed.
Results: The analysis of the findings showed that the prevalence of C. trachomatis infection
among young women 7.1%. Two-thirds (60 %) of infected C. trachomatis female students
had no specific clinical signs. Multivariable analysis showed that C. trachomatis infection
was twofold more common (OR – 2.37; CI 1.06; 5.3) among students who had specific STIs’
symptoms compared to those without such symptoms. The prevalence of C. trachomatis
infection was threefold higher among students who had 2–3 sexual partners compared to
those who had only one sexual partner (OR – 3.17; CI 1.03; 9.7). For the estimation of the
population prevalence, the costs of the present pooling strategy (177 pools) were reduced by
85 % compared with the analysis of all 533 samples. The costs of the present pooling strategy
with subsequent individual testing of all samples in each positive pool (177 pools and 87
individual samples from positive pools) for the estimation of the population prevalence as
well as the diagnosis of each individual sample were reduced by 70%.
Discussion: In order to save the reproductive health of young women, and money, in
Lithuania there could be a suggestion to politicians to implement new extensive screening
strategies, i.e. to organize centralized high-quality analysis of samples which can be taken
non-invasively; the detection of C. trachomatis should be performed in specialized regional
laboratories using pooled samples. Screening could be more cost-effective when risk factors
and specific clinical symptoms related to C. trachomatis infection are discovered.
Conclusions: the prevalence among young women was detected almost the same as in the
other European countries; pooling vaginal samples for nucleic acid detection of C. trachomatis
is an accurate and cost saving approach for screening to compare with testing of individual
samples.
CHLAMYDIA CONTROL
IN THE EUROPE
Marita van de Laar, European Centre for Disease Prevention and Control, Stockholm, Sweden
Background In 2007 a survey was carried out in EU/EFTA Member States with respect to
prevention and control of Chlamydia trachomatis infections. The report “Screening for
Chlamydia Review in Europe” outlines the basic requirements for guidelines, diagnostics,
case management, screening and surveillance. Guidance for Chlamydia control needed to
be developed with the purpose to lay down the scientific basis to support Member States
to strengthen and improve Chlamydia control. Target audiences are national programme
managers on sexual health and STI, policy makers at (inter)national level and experts in
the field.
37
ABSTRACTS
Objectives To present the latest surveillance data on Chlamydia and the ECDC guidance on
Chlamydia control in the Europe.
Methods A technical expert group on Chlamydia control in the Member States was
established. Comments from the ECDC’s Advisory Forum to the draft guidance were taken
into account. The 1990-2009 surveillance data have been collected in TESSy in 2010.
Results The guidance recommends a stepwise approach to Chlamydia control to ensure
that prevention and patient management are in place before complex interventions such
as screening are considered. A: Primary prevention (health promotion/education, school
programmes, condom distribution); B: Case management (routine case surveillance,
diagnostics, patient and partner management); C: Opportunistic testing (testing
routinely offered to specified group(s) of people attending clinical services); D: Screening
programme (organised provision of chlamydia testing to a substantial proportion of a
defined population). The first step to a comprehensive control programme is the adoption
of a national chlamydia control strategy based on consultation with key stakeholders.
The strategy should take into account the specific national opportunities and limitations
together with a review of the evidence. The strategy can be based on the stepwise approach
outlined in the ECDC guidance on Chlamydia control.
Nearly 344,000 cases of chlamydia have been reported in 2009; rates and trends will be
presented over two decades.
Conclusions The implementation of the Chlamydia guidance needs to be evaluated at both
national and the EU level. At the national level, programmes should monitor indicators
relating to the policies and guidelines of the programme, the implementation and processes,
and the outcome of the programme. At the European level, the target is to reduce the
proportion of countries reporting that no organised control activity exists.
THE ROLE OF CHLAMYDIA TRACHOMATIS ANTIBODY TESTING IN THE FIRST
SCREENING FOR TUBAL FACTOR INFERTILITY
Corresponding author: Jaroslavs Lakutins
Other authors: V.Fodina, J.Brikune
Institution of corresponding author: AVA Clinic, Vilandes 3, Riga, Latvia
Institutions of other authors: AVA Clinic, Vilandes 3, Riga, Latvia
Speciality: gynaecologist
Country: Latvia
ABSTRACTS
E-mail: jaroslav_lakutin@yahoo.com
Introduction: Chlamydia trachomatis (CT) in the female genital tract (FGT) usually remains
asymptomatic, undiagnosed and untreated. The main clinical form of CT infection is acute
cervicitis. CT may ascend to the upper FGT and can cause pelvic inflammatory disease (PID)
in up to 30% of cases. It increases the risk of tubal epithelial damage and tubal obstruction.
Therefore, an earlier CT infection can be the reason for tubal infertility (TI). The risk of TI after
the previous PID is estimated from 10 to 20%.
Objectives: The association between CT IgG antibodies, which still can be in the blood many
years after the treatment and tubal pathology, has been recognized before. Therefore, CT
38
antibody testing (CAT) can be used as a screening test for previous infection and possible
tubal pathology on the first visit to a gynecologist in the fertility clinic. The gynecologist
should separate patients with a high risk of tubal pathology for the next step of the
treatment from the patients with low risk.
Methods: The primary selection includes identification of the risk factors for tubal pathology
during the history taking, abnormalities during the ultrasound examination (USG) and
CAT. Laparoscopy is recommended for patients with known co-morbidities (PID, previous
ectopic pregnancy or endometriosis), and for patients with pathology after USG. In the case
of patients with no known co-morbidities and without abnormal USG finding, CAT helps to
detect women with low and high risk of tubal pathology.
Results: CAT has rather high negative predictive value (85-90%). It means, that the patients
with negative CT antibodies (low-risk patients) have low probability of tubal damage (less
than 15%). Performing hysterosalpingography (HSG) after CAT does not significantly change
the probability of tubal damage. However HSG as a method has a high specificity and can
very precisely confirm the absence of tubal pathology. Where appropriate expertise is
available, screening for tubal occlusion using hysterosalpingo-contrast-ultrasonography
(HyCoSo) should be considered because it is an effective alternative to HSG for women who
are not known to have co-morbidities.
Conclusions: Patients with positive CT antibodies (high-risk patients) have 53-60%
probability of tubal damage (positive predictive value). HSG recognise the tubal patology
with approximately the same probability. However, the risk of complications after HSG
for these patients is increased by up to 10%. Therefore HSG is relatively contra-indicated
for high-risk patients. Laparoscopy can be recommended to provide a final diagnosis.
CT infection should be excluded before operation for high-risk patients.
SHOULD WE ROUTINELY TEST MEN FOR CHLAMYDIA INFECTION IN ASSESMENT
OF THEIR FERTILITY POTENTIAL?
Corresponding author: Jaroslavs Lakutins
Other authors: O.Lakutina
Institution of corresponding author: AVA Clinic
Institutions of other authors: AURA-R Medical SIA
Speciality: gynaecologist
E-mail: jaroslav_lakutin@yahoo.com
Introduction: A history of genital tract infections (GTI) occurs in 1.6-10.3% of men attending
fertility clinics. Colonization of the male genital tract is probably common and usually
self-limiting. The male accessory sex glands often harbor microorganisms (also Ureaplasma
urealythicum and Chlamydia trachomatis (CT)), which may colonize the urogenital tract
without obvious sign of infection.
39
ABSTRACTS
Country: Latvia
Results: Semen is characterized by low bacteria counts and men without signs of infection
may have positive cultures. Occasionally, these microorganisms cause symptomatic
urethritis, prostatitis and epididymitis (male accessory gland infection (MAGI)) with
deterioration of the semen quality and leucocytospermia. Although CT is a frequent
pathogen and a well-documented source of MAGI, the infection does not seem to cause
obstruction of the reproductive tract in men, as it does in women. It is postulated that up
to 50% of men infected with CT are asymptomatic and in those with symptoms, the most
common presentation is urethritis.
Discussion: Studies suggest that most upper GTI in young men, also epididymitis, are
often attributable to CT. Epididymitis is thought to be important because fertility might
be affected due to inflammation, obstruction and functional impairment, especially where
both epididymis are affected. As well as creating a blockage to the movement of sperm, CT
may also cause epithelial damage that reduces spermatogenesis, induces immunological
reactions that hinder sperm, and reduces the female partner's fertility.
Conclusions: There are, however, no conclusive studies showing that men infected with
CT are less fertile than uninfected men. Male genital chlamydia infection is mainly a threat to
the female genital organs. Testing for Chlamydia trachomatis infection in a fertility clinic can
be recommended for men in the cases of symptoms of MAGI, leucocytospermia and in the
case of positive serological screening for CT in a female partner, if chlamydial infection was
not previously diagnosed.
CHLAMYDIA ANTIBODIES AS MARKERS FOR LATE COMPLICATIONS
IN COST-EFFECTIVENESS MODELS
Corresponding author: Jolande A. Land
Institution of corresponding author: Division of Reproductive Medicine, Department of
Obstetrics and Gynaecology, University Medical Center Groningen, University of Groningen,
Groningen, The Netherlands
Speciality: Obstetrics and Gynaecology
Country: The Netherlands
ABSTRACTS
E-mail: j.a.land@og.umcg.nl
Introduction: Chlamydia infections may give rise to pelvic inflammatory disease (PID) and
tubal infertility. The cost-effectiveness of chlamydia screening is determined by the rate
of complications prevented. There is a need for more precise estimates of PID and tubal
infertility to be inserted in economic models. Chlamydia IgG antibody testing (CAT) has
been introduced in fertility clinics as a screening test for tubal infertility. The feasibility of
CAT in screening programmes has not been evaluated.
Objectives: The aim of our research was to evaluate CAT as an intermediate marker for late
sequelae in a cost-effectiveness model. We estimated the prevalence of tubal infertility after
chlamydia infection based on data derived from the literature, and compared this with an
estimate based on a model including CAT.
40
Methods: We summarized the available literature dealing with the estimates for late
complications of chlamydia infections. We constructed a hypothetical model including CAT.
For the construction of the model we used data on CAT and tubal pathology in infertile
women, and data on seroconversion in PCR-test positive women.
Results: In the literature, the generally assumed risk of developing PID after lower genital
tract chlamydia infection varies considerably and is up to 30%. The estimated risk for
developing tubal infertility after PID is 10-20%. This implies that the risk of PCR-test positive
women developing tubal infertility would range between 0.1-6%. In our CAT-based model
the estimated a risk of tubal infertility is up to 4.6%.
Discussion: The prevalence of CAT positivity is high in infertile women with tubal pathology.
Therefore, CAT has been introduced as a screening test in fertility clinics. Based on the results
of our study, CAT might be usable in cost-effectiveness studies as an intermediate marker for
late sequelae of infection. Research questions that need to be addressed should deal with
the pathophysiology of antibody formation in chlamydia infections and the development of
tubal scarring.
Conclusions: The risk of developing tubal infertility after chlamydia infection appears low
(0.1-4.6%). Studies dealing with the transition from lower genital tract infection to tubal
infertility are needed to arrive at more precise estimates of the risk of late complications. In
these studies, antibody testing as a marker of a previous (asymptomatic) infection alongside
PCR testing should be evaluated to conclude whether CAT can be used as a surrogate marker
for adverse sequelae in screening programmes.
DIAGNOSTICS OF URINOGENITAL CHLAMIDIOSIS
COMMON FORMS
Corresponding author: Yuriy Vladimirovich Lobzin
Other authors: S.N.Sidorchuk, A.I.Poznyak, E.V.Boiko, V.S.Ageev
Institution of corresponding author: Medical Academy of Postgraduate Education
Institutions of other authors: Military-Medical Academy St.Petersburg
Speciality: infectious diseases specialist
Country: Russia
Introduction: Chronic urogenital chlamydia infection, causes a very wide range of sequelae associated with dissemination of C. trachomatis from the primary focus of infections to
sporadic organs. This leads to serious diseases such as genital or extragenital localization
(infertility, pregnancy failure, Chlamydia-induced reactive arthritis, oftalmohlamidioz,
perihepatitis, pelvic peritonitis). Hematogenous dissemination of agent is the basic unit in
the pathogenesis of common forms of chlamydial infection.
Objectives: Development of algorithms for diagnosis of chronic chlamydia extragenital
localization. 1002 patients at the age of 18-32 were observed and treated for acute urethritis,
41
ABSTRACTS
E-mail: sergei_sidorchuk@mail.ru
cervicitis (n=193), localized prostatitis, vesiculoprostatitis, adnexitis, salpingitis (n=340),
common Chlamydia trachomatis-induced reactive arthritis (n=274) forms of infection, acute
and chronic sinusitis associated with Chlamydia trachomatis (n=95), and meningitis and
encephalitis (n=100).
Methods: Several techniques were used: polymerase chain reaction (PCR), cell-culture, direct
immunofluorescence (DIM), enzyme multiplied immunoassay, indirect immunofluorescence,
and electron microscopy. Radioisotope scintigraphy and single-photon emission computed
tomography (SPECT) were also carried out.
Results: C. trachomatis has been found in the blood of 63.4 % of the patients with confirmed
chlamydial infection. Studying the diagnostic importance of detection of the pathogen in
blood has shown that in case of chronic genital infections and also extragenital pathologies,
the frequency of C. trachomatis detection in serum is 2-3.5 times higher, than in swab
material.
Discussion: Detection of C.trachomatis in the blood of patients, especially in cases of chronic
and complicated forms of chlamydia, has important diagnostic value, because it allows
using a fundamentally new approach of the direct detection of the pathogen, regardless
of the infection localization. Detection of hlamidemii can be used to diagnose chronic and
extragenital forms of chlamydial infection.
Conclusions: Using clinical material from the urinogenital tract cannot solve all the
diagnostics problems of different urinogenital chlamidiosis forms. Important clinical criterion
of common urinogenital chlamidiosis is the presence of chlamydia in blood. To elaborate
secondary foci of chlamidiosis it is advisable to use radioisotope scintigraphy.
WHAT DO WE REALLY KNOW ABOUT THE SPREAD OF CHLAMYDIA TRACHOMATIS
IN COMMUNITIES?
Corresponding author: Per-Anders Mårdh
Institution of corresponding author: Institute of Clinical Sciences, Department of Obstetrics
and Gynecology, Lund, Lund University, Sweden
Country: Sweden
E-mail: per-anders.mardh1@comhem.se
ABSTRACTS
In many countries, there is no surveillance system for genital chlamydial infections, reason
why the extent of carrier ship of Chlamydia trachomatis is unknown. This is then also true for
the extent of association of the agent with complications and sequelae that in other studies
have been established for Chlamydia trachomatis.
How is the situation in countries where genital infections by Chlamydia trachomatis are
monitored? The emerging of “the Swedish variant” highlighted that even in countries where
such infections are obligatory to report to authorities, the situation is still notably unclear.
In surveillance systems, there is generally no distinction between acute and likely chronic
infections. There is also no distinction made between the examined number of treated
and untreated cases. Likewise, those being tested as notified partners are not reported
separately.
42
The importance of understanding the prevalence of genital chlamydial infections in the
population to be tested is hardly ever highlighted.
In conclusion, a lot more is needed to sophisticate descriptive epidemiological surveys to be
able to serve any valuable analytic epidemiological research.
COMPETITION OF VACCINE AND NON-VACCINE HUMAN PAPILLOMAVIRUS TYPES
BEFORE AND AFTER MASS-VACCINATION
Corresponding author: Marko Merikukka
Other authors: Johanna Palmroth, Marjo Kaasila, Dan Apter, Jorma Paavonen, Matti
Lehtinen
Institution of corresponding author: National Institute for Health and Welfare, Oulu, Finland
Institutions of other authors: Department of Obstetrics and Gynecology, Kuopio University
Hospital, Kuopio, Finland, National Institute for Health and Welfare, Oulu, Finland The Family Federation of Finland, Helsinki, Finland, Department of Obstetrics and
Gynecology, University of Helsinki, Helsinki, Finland, School of Public Health, University
of Tampere, Tampere, Finland
Speciality: Statistics
Country: Finland
E-mail: marko.merikukka@thl.fi
Introduction: Replacement of multivalent vaccine covered serotypes of pneumococci by
non-vaccine serotypes is abolishing effectiveness of pneumococcal mass vaccination.
To understand the likelihood of type-replacement following vaccination against human
papillomavirus (HPV) types 16/18 we have studied competition of genital HPV types
by assessing multiple infections caused by seven most common genital HPV types
6,11,16,18,31,33 and 45 in fertile-aged Finnish females before and after mass-vaccination
between 1995 - 2004.
Methods: BEFORE mass-vaccination: First trimester serum samples from two consecutive
pregnancies (mean 2.5 years apart) were retrieved for a random 3 100 subsample of
123 000 women belonging to the Finnish Maternity Cohort. 42% had antibodies to at least
one HPV type at the baseline. AFTER mass-vaccination: In a sizeable phase III (PATRICIA) trial
sub-cohort of initially 4808 16-17 year-old Finnish women the HPV16/18 vaccine coverage
ranged between <1% and 20% by age-cohort and study community.
Results: BEFORE mass-vaccination: Highly significantly increased incidence rate ratios (IRR)
of seroconversion to another HPV type were consistently noted for HPV type 33 only, in both
HPV16 and HPV18 antibody-positive women: HPV16 antibodies; 16 and 33 antibodies
43
ABSTRACTS
Objectives: To explore type-replacement related to HPV mass vaccination using a prophylactic
HPV16/18 virus-like particle vaccine (CervarixT) with documented cross-protective
efficacy against HPV types 31/ 45, we assessed whether the IRR of non-vaccine HPV types
were significantly different in HPV16/ 18 vaccinated women as compared with hepatitis
A-vaccine recipients during a 36 month follow-up.
(IRR 3.2, 95% CI 2.0-5.2), and HPV18 antibodies; 18 and 33 antibodies (IRR 3.6, 95% CI 2.1-5.9);
irrespective of the presence of antibodies to other HPV types at baseline: HPV16 antibodies
only; 16 and 33 antibodies (IRR 2.9, 95% CI 1.6-5.4) and HPV18 antibodies only;18 and 33
antibodies (IRR 2.5, 95% CI 1.1-6.0). AFTER mass-vaccination: The IRR estimates of new HPV
types acquired during the follow-up in baseline HPV18 positive individuals as compared
to baseline HPV18 negative individuals did not differ statistically significantly from 1.0 for
non-vaccine, genital HPV types belonging to clades A5, A7, A9 or A10, neither among the
HPV16/18 vaccinated nor among the hepatitis A-vaccinated women. Discussion: BEFORE mass-vaccination: A competitive advantage for HPV33 over the other
genital HPV types before the era of HPV mass vaccination is suggested.
Conclusions: Our studies show no signs of type-replacement following HPV mass vaccination
even if some HPV types (HPV33) may have a competitive advantage at the population level in
the unvaccinated.
THE VALUE OF HOST GENETIC MARKERS COMBINED WITH CT SEROLOGY IN TUBAL
PATHOLOGY DIAGNOSIS
Corresponding authors: Sander Ouburg, Servaas A. Morré and the FP6 EpiGenChlamydia
Consortium
Institution of corresponding authors: Laboratory of Immunogenetics, Department of
Pathology, VU University Medical Center, Amsterdam, The Netherlands
Country: Netherlands
e-mail: s.ouburg@vumc.nl
ABSTRACTS
Chlamydia trachomatis (CT) infections are the world leading cause of preventable blindness
(trachoma) and the most prevalent sexually transmitted disease of bacterial origin which is
strongly associated with pelvic inflammatory disease, ectopic pregnancy, and tubal infertility.
Twin study-based findings of members of the FP6 EpiGenChlamydia Consortium identified
a 40% genetic predisposition to CT infections. The EpiGenChlamydia Consortium structured
transnational research to such degree that comparative genomics and genetic epidemiology
on large numbers of unrelated individuals can be performed to identify genetic markers to
be used for patient profiling.
Subfertility poses an enormous burden on healthcare and society throughout the world.
Worldwide, 15% of couples trying to conceive suffer from subfertility. Subfertility generally
described as a failure to conceive after one year of unprotected, regular sexual intercourse.
Approximately half of the couples suffering from subfertility will conceive spontaneously,
or after simple treatment. The other half needs more complex treatment, such as in vitro
fertilization (IVF) or other assisted reproduction procedures.
There are several causes of subfertility, which can be classified as ovulation disorders,
male factor subfertility, tubal damage, unexplained subfertility, and other causes, such
as endometriosis and fibroids. In women, one of the major causes of subfertility is tubal
pathology, with a prevalence of around 30%. In all of cases of tubal pathology, C. trachomatis
is the most common cause for infertility.
44
The reference standard for diagnosing tubal pathology in subfertile women is laparoscopy
performed usually after a CT IgG positive antibody test which is only positive in 50-60%
of women with tubal pathology. Despite the effectiveness, laparoscopy is costly, invasive
and has a low positive predictive value. Therefore there is a unmet medical need for better
diagnosing tubal pathology in women with subfertility, a need which could potentially be
met by adding genetic profiling to current diagnostic approaches. The current presentation
will discuss immunopathological mechanisms and host based genetic diagnostic
approaches.
PREVENTION OF CT AND HPV INFECTIONS:
ARE WE DOING ENOUGH?
Corresponding author: Jorma Paavonen
Institution of corresponding author: Department of Obstetrics and Gynecology, University
Hospital, Helsinki, Finland
Country: Finland
E-mail: jorma.paavonen@hus.fi
Chlamydia trachomatis (CT) infections of the genital tract are highly prevalent globally. CT is
a true obligate intracellular pathogen which has a unique growth cycle distinguished from
all other micro-organisms. Because of this slow growth cycle chlamydial infections tend to be
silent and chronic. The clinical spectrum of CT infections is wide. Most clinical manifestations
of CT genital infections are asymptomatic. Secondary prevention by screening for chlamydial
infections still remains important. CT vaccine development has proven difficult because
of the complex antigenic structure and limited knowledge of the protective antigens.
Guidelines or recommendations for annual screening of young sexually active individuals
for chlamydial genital infections have been implemented in many countries. However, the
effectiveness of screening programs has recently been challenged since population studies
suggest that screening programs have been strikingly ineffective in lowering chlamydia
rates. By contrast, the rates of reported chlamydial infections have been increasing in many
countries regardless of implementation of screening.
Human papillomavirus (HPV) infection is the most common viral sexually transmitted
infection. HPV infections are extremely common, and most young adults are exposed to
HPV soon after sexual debut. Although most HPV infections resolve, persistent infection
by one or more of the oncogenic HPV types can cause cervical neoplasia. HPV can also
cause other anogenital neoplasias and a smaller proportion of oropharyngeal neoplasias.
Worldwide, cervical cancer is the second most common cancer in women and a major
cause of cancer death in women worldwide. Secondary prevention by cytologic screening
has been effective in some countries, but the screening policies vary widely. Therefore, the
incidence rates of cervical cancer differ strikingly between countries, also within Europe. Due
to mass screening, the disease burden has shifted to management of cervical intraepithelial
45
ABSTRACTS
Thus, there is major frustration with opportunistic or organised screening programs,
implementation of management guidelines and contact tracing efforts. Screening activity
may certainly benefit individuals but seems to have little public health impact.
neoplasias (CIN). This causes health problems and drains health care resources. The
development of prophylactic HPV vaccines has been a remarkable success story. Primary
prevention by these vaccines can substantially reduce the public health and economic
burden of cervical precancer and cancer and other HPV-associated diseases. Clinical efficacy
trials have demonstrated that these vaccines are strikingly effective in the prevention of high
grade cervical, vaginal and vulvar precancers. These trials have also raised several questions
and challenges to be addressed to assess the public health impact on all HPV-associated
diseases, and to design the most effective vaccination strategy. These include duration
of immune response, vaccination of males, efficacy against other HPV-related cancers
bewyond cervical cancer, postmarketing safety surveillance, impact of vaccination on
existing screening programs, potential HPV type-replacement after widespread vaccination,
and finally feasibility of HPV vaccination in developing countries where the disease burden
remains enormous.
Although major improvement has taken place in recent years in our armamentarium to
prevent sexually transmitted CT and HPV infections the question still remains: Are we doing
enough?
EPIDEMIOLOGICAL SURVEILLANCE OF CHLAMYDIA TRACHOMATIS
INFECTION IN LATVIA
Corresponding author: Jurijs Perevoscikovs
Other authors: Violeta Mavcutko
Institution of corresponding author: Epidemiological Safety and Public Health Department,
State Agency "Infectology Center of Latvia"
Institutions of other authors: HIV/ AIDS Surveillance and Prevention Unit, Epidemiological
Safety and Public Health Department, State Agency "Infectology Center of Latvia"
Speciality: epidemiology
Country: Latvia
ABSTRACTS
E-mail: jurijs.perevoscikovs@lic.gov.lv
Introduction: Chlamydia trachomatis is the most commonly reported sexually transmitted
infection (STI) in Latvia. The proportion of Chlamydia infection increased in STI structure
up to 66% last year. Genital Chlamydia infection is a significant public health concern
because untreated infections cause complications with serious consequences on women's
reproductive health. There are four STI under mandatory notification in Latvia. STI are in the
4th place in the structure of the notifiable infections in Latvia.
Objectives: Epidemiological surveillance system monitors the spread of Chlamydia
trachomatis and provides important information for effective control of the infection.
Chlamydia is easily treated with antibiotics and a preventable disease (safe sex, condom use).
An important aspect of prevention involves the evaluation of sexual partners to prevent
re-infection and further spread of the disease.
46
Methods: C. trachomatis is a mandatory notifiable infection in Latvia. Data from the national
computer-based surveillance system VISUMS were used to describe epidemiological trends
for the period from 2008 to 2010. The case definitions used in Latvia are based on the
European Union case definitions. The reports of Chlamydia cases to the national surveillance
system contain individual laboratory notification from a diagnostic laboratory and an
individual clinical notification from health care providers.
Results: In Latvia, altogether 2873 Chlamydia cases (58.5% of all cases of STI) were reported
to the national surveillance system from 2008 to 2010. The proportion of Chlamydia in STI
structure increased from 46% in 2008 to 66% in 2010. Up to 66% of all cases were reported
in the age group 18 - 29 years. The proportion of females in structure of Chlamydia infection
increased from 46% in 2008 to 64% in 2010. In women of age group 18-29 years the
incidence rate of genital Chlamydia increased by 2,2 times, from 105,1/100 000 in 2008 to
238,4/100 000 in 2010. During the period of 2009 to 2010 reported cases in pregnant women
increased from 57 (9.4% of all reported females cases) to 174 (26%) cases.
Discussion: In Latvia the highest incidence rate of Chlamydia infection was found in young
women of reproductive age. Chlamydia is usually asymptomatic in both women and men.
In women, it may result with complications, which is a major cause of tubal infertility, ectopic
pregnancy. The number of diagnosed and treated cases depends on the number of persons
tested. In Latvia legislation obliges testing for asymptomatic cases only for pregnant women,
but there is a lack of screening program for populations.
Conclusions: During the last two years the number of notified Chlamydia cases in females
notably increased (by 206%). It could be explained by the following reasons: since July 2008
all microbiology laboratories were involved in notification system and significant effort has
been made to improve the completeness of reporting by health care providers in Latvia. The
development and implementation of a screening programme is very important for further
improvement of surveillance and control of Chlamydia infection.
GENOTYPING OF CHLAMYDIA TRACHOMATIS
STRAINS
Corresponding author: Mirja Puolakkainen
Other authors: Suvi Niemi and Eija Hiltunen-Back
Institutions of other authors: Haartman Institute, Department of Virology, University of
Helsinki, and HUSLAB, Department of Virology and Immunology, Helsinki University Central
Hospital, Clinic of Venereal Diseases, Skin and Allergy Hospital, Helsinki University Central
Hospital, Finland; National Institute of Health and Welfare, Helsinki, Finland; Helsinki, Finland
Speciality: clinical microbiology
Country: Finland
E-mail: mirja.puolakkainen@helsinki.fi
47
ABSTRACTS
Institution of corresponding author: Haartman Institute, Department of Virology, University
of Helsinki, and HUSLAB, Department of Virology and Immunology, Helsinki University
Central Hospital, Helsinki, Finland
Introduction: Since 1995, the number of Chlamydia trachomatis infections has been officially
notified in Finland. Lately, there has been around 14 000 notified cases annually (250 cases/
100 000 inhabitants). C. trachomatis infections are diagnosed with sensitive and specific nucleic
acid amplification tests (NAATs), but these tests do not differentiate between genotypes.
We wanted to further refine the current laboratory diagnosis of C. trachomatis infections.
Objectives: Our purpose was to study molecular epidemiology of urogenital and extragenital
C. trachomatis strains, and to study the occurrence of the Swedish new variant of C. trachomatis
in Finland.
Methods: We set up real-time quantitative PCR based methods for genotyping C. trachomatis
types D-K (ompA-PCR) and L1-L3 (pmpH-PCR) as well as the Swedish plasmid variant of
C. trachomatis (plasmid-PCR).
Results: Of the 160 C. trachomatis positive urogenital samples analyzed, 90% could be
genotyped with the ompA real-time PCR. The three most prevalent genotypes were E, F and
G. No L types were detected. Although the genotype E was the most common genotype
among C. trachomatis strains, the Swedish new variant was rarely found (only 2/469). Of
the 100 C. trachomatis strains detected in extragenital specimens, 79 could be genotyped.
Five rectal swab specimens harbored the genotype L2b. All these patients belonged to the
identified risk groups.
Discussion: Typing of C. trachomatis strains can be used in epidemiologic surveillance, to
assess the changes in genotype distribution and to reveal transmission patterns in sexual
networks. This information is of value in efforts to monitor and prevent spread of
C. trachomatis infections.
Conclusions: The most prevalent genotype in Finland was E (40%). The Swedish variant was
rare in Finland despite our close proximity to Sweden. Also in Finland, L2b genotypes were
detected in rectal swabs taken from HIV positive MSM.
NEW METHODS FOR CERVICAL CANCER DETECTION IN LITHUANIA. VALUE OF
HIGH-RISK HPV-DNA TESTING IN THE TRIAGE OF WOMEN WITH ASC CATEGORY
FROM LIQUID BASED CYTOLOGY
Corresponding author: Jolita Rimiene
Institution of corresponding author: Diagnostics Pathology, Vilnius University
Country: Lithuania
ABSTRACTS
E-mail: jolitarimiene@patologija.lt
Background: Cervical cancer in Lithuania is the fourth commonest cancer among women
with malignancies. Since 1992 Lithuania have the increasing rates in incidence of cervical
cancer among Europe countries and the highest in Baltic sea region countries. The
Lithuanian ministry of Health started nation- wide cervical cancer screening program in the
middle of 2004. During the program overall was diagnosed 7 778 HSIL and 323 cancer cases.
In the screening group during program period we observed statistically significant increase
in incidence of stage I cervical cancer and decrease in stage II. The changes in incidence
of stages III and IV cervical carcinoma we did not observed. During the program period we
observed statistically significant increased incidence of CIN3/CIS – from 1,7/100 000 in 1999
to 31,1/100 000 in 2008. The incidence was increasing by 34,4% in all age group and by
48
33,7% in 25–59 age group and by 26,4% in 60 and older years age group.
Cytological screening has reduced the incidence of invasive cervical cancer, but as a result
of screening, many women are diagnosed as having equivocal cytological abnormalities (eg,
atypical squamous cells, herein referred to as ASC). 11 pathology laboratories are accredited
to perform conventional Pap tests. According Screening Program period in Lithuania
was diagnosed 16 545 (3%) ASC cases, high number of unsatisfactory smears (2,3-9,6%).
Many laboratories are performing HPV testing from fresh cytological material after the
conventional Pap smear. We introduce Cobas® 480 HPV detection system in combination
with ThinPrepTM liquid based cytology. We analyzed the initial data from the first 1397
ThinPrepTM Pap tests. 13.7% abnormal cytology cases were found. Low 0.6% unsatisfactory
and 1,1% ASC ratio was observed. A high number of LSIL and HSIL (8% and 4.6%) cases was
detected. Endocervical adenocarcinoma case was diagnosed. The difference number of SIL
cases was detected in healthy women group and women with risc factors: 5.7% LSIL and
2.5% HSIL, 7% LSIL and 7% HSIL respectively. HPV infection was found in 39 (73.6%) of 53
tested HSIL and 14 (77.8%) of 18 LSIL ThinPrepTM vials. HPV was detected in 8 (47%) of 17
ASC cases. In 5 ASC-H and 3 ASC-US cases HR-HPV types was detected. 4 of 5 women who
underwent conisation CIN2/CIN3 lesion was found in the histological material.
Conclusions. Liquid based cytology and reflex HPV DNA testing is effective measure for early
cervical cancer detection. HPV-DNA testing may serve as a means to better select which
patients with ASC-H and ASC-US on Pap cytology should undergo colposcopic examination
and conisation. This approach potentially may reduce medical costs and eliminate the need for
routine repeat cytology, colposcopic examination among patients with ASC-US Pap smears.
DO ABBOTT REALTIME CT AND -HIGH RISK HPV MEET THE REQUIREMENTS FOR
LABORATORY SCREENING AND DIAGNOSIS OF CHLAMYDIA TRACHOMATIS AND
HUMAN PAPILLOMA VIRUSES?
Corresponding author: Edwin Roovers
Institution of corresponding author: Abbott Molecular
Speciality: Marketing Manager EMEA&India
Country: The Netherlands
Introduction: Various forms of Chlamydia screening using NAAT have been suggested but
they were implemented only in a limited number of countries. HPV DNA testing has become
an important part of cervical carcinoma screening and management algorithms in several
countries. Clinical trials were conducted to show the performance of the RealTime CT and
RealTime High Risk HPV assays as well as how they compare versus other methods. Also
more practical criteria for the laboratory will be discussed.
Methods: The m2000 system is a fully automated IVD system for nucleic acid extraction,
real-time PCR and results reporting. A multicenter clinical study was conducted to evaluate
the performance characteristics of the Abbott Real-Time CT/NG assay compared to Aptima
Combo 2 Assay (Gen-Probe), ProbeTec ET CT/GC (BD). For HPV the clinical performance
of the Abbott Real-Time High Risk HPV test and Hybrid Capture 2 HPV DNA Test was
prospectively compared in a screening setting.
49
ABSTRACTS
E-mail: Edwin.Roovers@abbott.com
Conclusions: The Abbott m2000 provides laboratories with ease of use, reliability and
efficiency for a broad spectrum of IVD assays. It is also an open system for running in house
methods. Taking factors into consideration such as assay sensitivity & specificity, accuracy,
efficiency and reliability it can be confirmed that Real-Time CT and Real-Time High Risk
HPV do meet the high standard requirements for laboratory screening and diagnosis of
Chlamydia trachomatis and human papillomaviruses.
IL6 -174 G>C POLYMORPHISM, HPV INFECTION AND CERVICAL INTRAEPITHELIAL
NEOPLASIA
Corresponding author: Iona Rotar1
Other authors: Daniel MURESAN1, Radu POPP2, Sorana D. BOLBOACA3, Felicia PETRISOR2,
Cristina BUTUZA4, Silvana APOSTOL4, Georgiana COROIU5, Florin V. STAMATIAN1
Institutions of other authors: 1- „Iuliu Hatieganu” University of Medicine and Pharmacy
Cluj-Napoca, 1st Department of Obstetrics and Gynecology, 3-5 Clinicilor,
400006 Cluj-Napoca, Romania
Country: Romania
E-mail: cristina.rotar@umfcluj.ro
Introduction: The relationship between chronic inflammation and cancer is well-know
especially for HPV and cervical cancer. Interleukin-6 (IL-6) an inflammatory cytokine can plays
either inhibiting or stimulating roles in carcinogenesis.
Objective: In this study we aim to analyze the relation between cervical cancer and IL6 -174
G>C polymorphism.
Materials and methods: A case control study has been conceived. The cases were represented
by 128 patients diagnosed with cervical intraepithelial neoplasia, positive at HPV HR testing,
while 111 controls, negative for intraepithelial neoplasia and negative at HPV HR testing were
included in the control group. For each patient a physical examination has been performed.
Both cervical and peripheral blood sample were obtained. From the cervical probe cytology
and HPV HR testing has been done, while DNA has been extracted from blood. Later IL6 -174
genotyping was detected using a PCR-RFLP technique.
ABSTRACTS
Results: The absolute genotypes frequencies were for cases: G/G - 60, C/G – 52, C/C- 16
respectively for controls G/G – 19, G/C- 47, C/C – 19. Chi square had a value of χ2 = 0.968;
df = 1; p = 0.325 while considering G/C&/C/C genotype as a risk factor, respectively χ2 = 1.014;
df = 1; p = 0.314 when only homozygous C/C genotype was taking into consideration as a
risk factor. OR had values of 1.2, CI 95% [0.774, 2.163] for G/C&/C/C genotype, respectively
OR-1.446, CI 95% [0.704, 2.970] for T/T genotype considered as a risk factor.
Discussion: HPV represents a mandatory condition for cervical carcinogenesis. In particular
the genetic polymorphism at -174 could be followed by a change in IL-6 local and serum
level. By the binding of the latter interleukin to its receptor a Jak pathway is activated. The
implication of Jak kinase has been previously reported especially for hematologic cancer but
also for solid carcinomas.
Conclusion: The presence of at least one variant allele could be considered as a risk factor for
cervical intraepithelial neoplasia but the association is weak.
50
WHAT SHOULD BE THE BASIS OF A NEW CERVICAL
SCREENING PROGRAMME?
Corresponding author: Peter Sasieni
Country: United Kingdom
E-mail: p.taylor@qmul.ac.uk
Cervical cancer rates have been greatly reduced in many industrialized countries through
screening programmes based on cervical cytology. Unfortunately this technology has had
little impact in low and middle income countries. Today HPV testing and visual inspection
methods are available as alternatives to cytology. Additionally, HPV vaccination offers a
one-off approach to cervical cancer prevention. I will consider the pros and cons of different
approaches to screening and argue that except in places with an established and effective
cytology based screening programme, screening should be based on HPV testing with other
tests reserved for triage of HPV positive women. Whilst vaccination may be seen as the
solution for the future, a need for good screening will remain for many years to come.
HPV SURVEILLANCE
IN THE VACCINE ERA
Corresponding author: EJ Savage
Other authors: R Howell-Jones, K Soldan
Institution of corresponding author: Health Protection Agency
Specialty: STI surveillance
Country: United Kingdom
Introduction: An important component of any vaccination programme is monitoring
to assess the impact of the programme. The Human papillomavirus (HPV) vaccine was
introduced into the UK immunisation programme in 2008 and is offered to all females aged
12-13 years of age and a “catch-up” campaign was conducted to reach those aged 14 to
18 years. High levels of uptake have been achieved with 76.4% of 12-13 year old females
completing the three dose course in 2009/10. Cervarix, the vaccine currently used in the
UK programme, protects against HPV types 16 and 18 which are associated with over 70%
of cervical cancers in the UK.
Conclusions: We will present an overview of HPV surveillance programmes that have been
established to monitor key outcomes with a particular focus on England. As well as vaccine
safety and coverage, systems are now in place to monitor changes in the frequency of
vaccine (and non-vaccine) HPV types, vaccine effectiveness, immunogenicity and
herd-immunity.
51
ABSTRACTS
Email: emma.savage@hpa.org.uk
THE AMAZING STORY OF THE SWEDISH NEW VARIANT
OF CHLAMYDIA TRACHOMATIS (nvCT)
Corresponding author: Magnus Unemo
Institution of corresponding author: National Reference Laboratory for Pathogenic Neisseria,
Department of Laboratory Medicine, Microbiology, Örebro University Hospital, Örebro,
Sweden
Speciality: Microbiology and Molecular Biology
Country: Sweden
Email: magnus.unemo@orebroll.se
Introduction: In late-2006, the new variant of Chlamydia trachomatis (nvCT) was reported
in Sweden. The nvCT contains a 377 bp deletion within its plasmid, which encompasses
the targets in the commercial diagnostic PCR systems from Roche and Abbott at the time.
The nvCT rapidly spread nationwide in Sweden, undiagnosed by these systems, and caused
thousands of false negative reports.
Objective: To provide a comprehensive overview of all published data regarding nvCT, its
emergence, epidemiology, phenotypic and genetic characteristics, and effects on public
health and C. trachomatis surveillance.
ABSTRACTS
Overview: The nvCT is clonal and has been assigned serovar/genovar E, multilocus sequence
typing [MLST] ST 21 (hctB), 19 (CT058), 1 (CT144), 2 (CT172), and 1 (pbpB), as well as variable
number of tandem repeats [VNTR] type 8.7.1 (CT1335, CT1299, and CT1291, respectively).
Genome sequencing and detailed phenotypic characterisation have shown that the
biological fitness of nvCT in vitro, when compared to wild type CT strains, is unaltered. This
is supported by epidemiological and most clinical observations. Accordingly, the initial
rapid transmission of nvCT in Sweden was due to the diagnostic selective advantage and
its introduction into a high-frequency transmitting population. The proportions of nvCT
cases now appear to be converging towards equilibrium with the wild type CT strains in
the different Swedish counties. The absence of any isolates from before 2000 and statistical
estimations suggest that nvCT emerged 4–6 years before it was detected. Interestingly, the
nvCT remains rarely reported beyond the Nordic countries and the epidemiological reasons
for this finding are not elucidated.
Summary: The emergence of nvCT in Sweden created a unique opportunity to study the
emergence, spread and effects on public health of a single strain within both the human and
bacterial populations. The nvCT had a substantial impact on C. trachomatis identification,
epidemiology, and public health in Sweden. The lessons learned, which are also applicable
to other infectious diseases, include; importance of epidemiological surveillance and
especially detailed investigation of changes in the incidence and epidemiology of infection,
the frequent participation in appropriate quality assurance schemes and the careful design
of diagnostic assays (ideally with multiple targets). It is crucial to have diverse diagnostic
systems and response plans prepared for similar situations in the future.
52
FIRST SCREENING PROJECT REGARDING CHLAMYDIA TRACHOMATIS INFECTION
IN ALBANIA
Corresponding author: Adela Vasili
Other authors: Lila SHUNDI, Brunilda VILA, Silva BINO, Eugena ERINDI, Gjergji KOJA,
Dritan ULQINAKU, Shpetim QYRA, Mimoza BASHO
Institution of corresponding author: Institute of Public Health, Tirana, Albania
Institutions of other authors: Institute of Public Health, Tirana, Albania
Speciality: Epidemiologist
Country: Albania
E-mail: adelavasili@yahoo.com
Introduction: Chlamydia trachomatis infections are now recognized as the second most
frequent cause of sexually transmitted diseases (STD) worldwide. Considering that in
Albania, till now, a national control program on genital chlamydial infection has not been
implemented, the development of a screening project in Albania was necessary.
Objective: The aim of this study was to determine the prevalence of Chlamydia trachomatis
genital infections, as well as the feasibility of molecular screening in Albanian women of
reproductive age.
Materials and Method. Study population: From October to December 2010, endocervical swab
specimens were collected from 241 sexually active women, under 40 years and consulting in
women’s health services in 4 districts Tirana, Durres, Vlore and Fier. The swabs were collected
using Amplicor STD Swab Specimen Collection and Transport Kit (Roche Diagnostics Systems).
The selected women filled in a standardized questionnaire. Molecular amplification analysis
was performed on all samples using Cobas AmplicorTM CT Test (Roche Diagnostics Systems) on
the COBAS Amplicor instrument, following the manufacturer’s instructions.
Conclusion: This study showed the prevalence and revealed the risk factors related to
C. trachomatis infections in Albania. This study, being the first of this kind in Albania, used
a nucleic acid amplification test for the identification C. trachomatis in a selected population.
This study creates the opportunity for implementing a national control and prevention
program on genital Chlamydial infection, which does not exist so far.
Additional information: In Albania there is no data on the incidence of chlamydial infection,
which is not mandatorily to be reported by the laboratory or clinic. In Albania there was a
national program to control and prevent HIV-AIDS, but for chlamydial infections, this is the
first initiative.
53
ABSTRACTS
Results: A total of 241 women of median age 30 years (range 17-40 years) were studied. The
prevalence rate was 7.5%, (95% CI=4.5%-11.5%). The prevalence differed significantly by
age: less than 20 years the prevalence is 0; 11.9% (5/42), among women 20-24 years, 9.1%
(6/66) for the women 25-29 years and 5.8% (7/121) among those over 30 years. A new sexual
partner in the last 6 months was the main risk factor to be C. trachomatis infected with an OR
7.2 (95% CI=2.38-23.2). Neither reason for consulting nor the existences of symptoms were
significantly associated with being C. trachomatis infected.
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RAPID DIAGNOSTICS
FOR CHLAMYDIA TRACHOMATIS
Corresponding author: Abhay Vats
Other authors: Ashok Gurung, Ashish Yeri, Di Gao, David Mead, Toni Darville, Shiela West,
Ishwad Chandra
Institution of corresponding author: University of Pittsburgh
Institutions of other authors: Johns Hopkins University, Lucigen Corp
Speciality: Pediatrics
Country: USA
E-mail: abhsy.vats@chp.edu
Introduction: Chlamydia infections are a major cause of morbidity and preventable blindness
in the developing world. In addition, they represent a major cause of sexually transmitted
diseases worldwide, and many such cases can be difficult to diagnose due to lack of
appropriate testing facilities in poor resource settings.
Objectives: We aimed to develop a low cost detection method for Chlamydia trachomatis,
which would be useful for both eye and genital infections. It is based on Loop mediated
isothermal DNA amplification technique (LAMP).
Methods: We developed primers and amplification protocols for several targets in the
Chlamydia genome as well as a lateral flow detection method for amplicons. Primers were
labeled with FITC, Biotin and Digoxigenin for detection in the lateral flow strips. A self
contained cassette was used for detection of the LAMP amplicons.
Results: The method was able to successfully amplify Chlamydia DNA using a low cost heater
(heater cost <$2.00). The amplicons were detectable within 5 minutes after the reaction on
the lateral flow strips. The whole procedure takes less than 30 minutes and cost less than
$5.00 per reaction with sensitivity and specificity similar to PCR.
Discussion: We have developed a low cost and highly sensitive and specific amplification and
detection system for Chlamydia trachomatis diagnostics.
ABSTRACTS
Conclusions: Isothermal DNA amplification and lateral flow detection methods could be
useful for managing this infection in low resource settings. Further development and clinical
testing of the assay is being planned.
56
EPIDEMIOLOGY AND CONTROL OF GENITAL CHLAMYDIA TRACHOMATIS
INFECTION IN SWEDEN
Corresponding author: Inga Velicko1,2
Other authors: Kühlmann-Berenzon S1
Institution of corresponding author: 1- Department of Analysis and Prevention, Swedish
Institute for Communicable Disease Control, Solna, Sweden
2- Department of Medical Epidemiology and Biostatistics, Karolinska Institutet, Stockholm,
Sweden
Institutions of other authors: 1- Department of Analysis and Prevention, Swedish Institute
for Communicable Disease Control, Solna, Sweden
Speciality: Epidemiologist
Country: Sweden
E-mail: inga.velicko@smi.se
ABSTRACTS
Genital Chlamydia infection is mandatory notifiable in Sweden through the coded,
anonymous electronic web-based notification. The majority (86%) of all reported cases is
between the ages of 15 and 29 years, and 57% are females. Chlamydia incidence increased
significantly between 1997 and 2004, from 157 to 356 cases per 100 000 inhabitants. The
following emergence of a new genetic variant of Chlamydia trachomatis in 2006, distorted
the “natural” history of Chlamydia epidemiology as cases were missed due to the inability of
some of the diagnostic-kits to detect this new variant. The catch-up of cases that followed
imitated an artificial increase in incidence that was clearly observed in 2007 and 2008.
Chlamydia incidence is significantly dependant on testing activity, which has increased
in Sweden continuously since 1997. In 2009 more than half a million persons were tested,
and 7% of those were positive. However, only one third of tested persons were males.
Given the figures and the limited testing coverage, it is safe to say that Sweden does not
yet have an effective control on the spread of the disease. For this reason, and based on the
epidemiologic background and data from past surveys on sexual health, the authorities
initiated a “National Programme for chlamydia prevention with focus on adolescents and
young adults, 2009-2014”. The programme’s measures are expected to show an effect
after some years and should therefore be critically and continuously evaluated in order to
incorporate adjustments as needed.
57
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58
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SELF-SAMPLE HPV-TESTS - EFFECTS IN CERVICAL CANCER SCREENING ATTENDANCE
AND COVERAGE IN FINLAND
Corresponding author: Anni Virtanen
Other authors: Pekka Nieminen, Tapio Luostarinen, Ahti Anttila
Institution of corresponding author: Mass Screening Registry, Finnish Cancer Registry
Institutions of other authors: Department of Obstetrics and Gynaecology, Helsinki University
Central Hospital, Jorvi Hospital Mass Screening Registry, Finnish Cancer Registry
Speciality: cervical cancer screening
Country: Finland
E-mail: anni.virtanen@cancer.fi
Introduction: Attendance in screening is an important determinant of cervical cancer. In Finland,
the overall attendance rate in the organized screening programme is 70% and attendance among
women aged 30-35 only 50-60%. High-risk human papillomavirus (hrHPV) DNA detection based
testing on patient obtained samples (self-sampling tests) are sensitive in finding cervical cancer and
its precursors. Previous experience also suggests a good effect among non-attendees of screening.
Objectives: We assessed the effects of self-sampling among the non-attendees of the routine
screening programme of a Finnish municipality. Main outcomes were increases in screening
attendance and coverage (coverage of any Pap-smear within the 5-year screening interval)
achieved by self-sampling kits, reminder letters or the combination of these two. Additional
outcome was to compare the prevalence of hrHPV test-positivity among first invitation and
second/third intervention attendees.
Methods: Non-attendees after the primary invitation were randomised to receive either a
self-sampling kit (2,397 women) or a reminder letter (6,302 women). One fourth (1,320
women) of arm non-attendees after a reminder letter also received a self-sampling kit as a
third intervention. Effects on screening coverage were assessed according to the self-reported
previous Pap smear history of the participants.
Discussion: Our results suggest that if self-sampling serves as the third intervention after two
reminder letters, on a national scale an increase in participation rate of organized screening is
achievable from 70 to 80% or more. Self-obtained samples are most likely more often HPV-positive
because they also reveal vaginal or even vulvar HPV infections in addition to the cervical ones.
Conclusions: In increasing the attendance at organized screening, self-sampling can feasibly
be used as an option for a reminder letter, or as an addition to it. More research is needed
to assess effects in different settings, as well as with regard to prevented cancers and
cost-effectiveness.
59
ABSTRACTS
Results: Participation rate by self-sampling, 31.5% (CI: 29.7, 33.4%), was significantly higher than
with a reminder letter, 25.9% (CI: 24.8, 26.9%). The adjusted relative risk for participation by
self-sampling was 1.21 (CI 1.13-1.3). Total attendance increased from 64.9 to 76.0% with
self-sampling and from 64.6 to 73.8% with a reminder letter. Combining the interventions reached
40% of non-attendees and increased total attendance from 63% to 78%. Only app. 20% of the
participants in all three groups increased screening coverage (previous Pap-smear ≥5 years ago or
never). Self-obtained samples were more often HPV-positive than provider obtained ones, 12-13%
versus 7-8%.
Abbott RealTime
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• More confidence in detection of disease compared to cytology*
• HPV-typing results will help guide clinical management decisions**
Flexible Sample Collection Options for
RealTime High Risk HPV on m2000 system
• Validated with ThinPrep PreservCyt Solution
• Validated with SurePath Preservative Fluid
(pre/post-gradient)
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* Huang et al. Journal of Clinical Virology, 2009; 45 (Suppl 1): 19 – 23
** Kahn et al. Journal of the National Cancer Institute, 2005; 97 (14):1072 – 1079
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Addressing Chlamydia
testing needs
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