SYNLAB Budapest Diagnostic Center GenoID Molecular Diagnostic Laboratory

READY FOR ASSISTANCE SYNLAB Budapest Diagnostic Center GenoID Molecular Diagnostic Laboratory Human Papillomavirus (HPV) testing Sexually Transmitted...
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READY FOR ASSISTANCE

SYNLAB Budapest Diagnostic Center GenoID Molecular Diagnostic Laboratory Human Papillomavirus (HPV) testing Sexually Transmitted Infections (STI) panel

About us SYNLAB Budapest Diagnostic Center GenoID Molecular Diagnostic Laboratory (GenoID) provides high-throughput, fast and accurate PCR-based pathogen detection as a referral laboratory for institutes all over the world for diagnostics and research. GenoID Laboratory is one of the most highly automated PCR labs in Europe. The requirement of cost efficient operation and the ambition for constant improvement in automation has resulted in remarkable know-how in operating a PCR laboratory. Over the past years the laboratory has performed more than half a million PCR tests including over 300,000 HPV tests for clients in Croatia, Hungary, Georgia, Romania, Serbia, Sweden, Turkey, UAE, UK, etc. Our services concentrate on two large areas: molecular microbiological diagnostic testing and human molecular genetic testing. Our laboratory is certified to ISO 9001:2001 and ISO 13485:2003 standards. We hope we can shortly welcome you among the growing number of European gynaecologists, oncologists, dermatologists and urologists who seek high-quality and cost-efficient HPV typing, STI and genetic diagnostic service.

Number of GenoID PCR tests in 2015

Mycoplasma hominis Treponema pallidum Atopobium vaginae Gardnerella vaginalis HIV1/2-HCV-HBV Trichomonas vaginalis Herpes simplex virus 1,2 Molecular genetics Neisseria gonorrhoeae Mycoplasma genitalium Ureaplasma urealyticum Chlamydia trachomatis Human papilloma virus

142 858 1680 3688 4367 4868 7167 15315 21448 22766 25654 35866 45845 0

10000

20000

30000

40000

50000

Contact us for more information: www.genoid.net [email protected] Tel.: 00 36 1 465 01 24

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HPV detection and genotyping Human Papillomavirus (HPV) testing It is well known that the presence of HPV shows strong correlation with the incidence of cervical cancer and its pre-cancerous states. HPV can be detected in almost 100% of cervical carcinoma patients. HPV genotypes are classified as low-risk or high-risk in regard to their ability to cause cervical carcinoma. The cumulative lifetime incidence of HPV infection is 70-80 percent in many countries. A certain ratio of women who become infected with HPV develop a persistent infection that can lead to precancerous changes in the cervix. The traditional screening method (PAP smear) has a shortcoming in that women with negative PAP smear test results can continue to develop cervical carcinoma. The interest is growing worldwide in the use of HPV DNA test to supplement cervical cancer prevention PAP smear programs. Combining these two methods can help eradicate cervical cancer. Furthermore, HPV DNA testing is the optimal way to determine the treatment after borderline PAP smear test results such as ASCUS (atypical PAP or atypical squamous cells of undetermined significance). Several large population-based trials have demonstrated that tests detecting human papillomavirus (HPV) DNA of oncogenic types (hrHPV) are more effective in reducing cervical cancer than the cytology. [1, 2]. The latest cervical cancer screening guidelines recommend hrHPV DNA testing as a primary screening test [3, 4]. The meta-analysis by Arbyn et al. demonstrated that validated PCR-based tests detecting hrHPV DNA are as accurate on self-samples as on clinician-collected samples [5]. According to the latest cervical cancer screening guideline offering self-sampling kits for HPV testing is an opportunity to reach women who do not attend the regular screening program, thereby increasing the effectiveness of the overall program [3, 6].

References 1. Ronco, G., et al., Efficacy of HPV-based screening for prevention of invasive cervical cancer: follow-up of four European randomised controlled trials. Lancet, 2014. 383(9916): p. 524-32. 2. Dillner, J., et al., Long term predictive values of cytology and human papillomavirus testing in cervical cancer screening: joint European cohort study. BMJ, 2008. 337: p. a1754. 3. Karsa, L., M. Arbyn, and H. Vuyst, European guidelines for quality assurance in cervical cancer screening. Summary of the upplements on HPV screening and vaccination. Papillomavirus research, 2015. 1: p. 22-31. 4. Huh, W.K., et al., Use of primary high-risk human papillomavirus testing for cervical cancer screening: interim clinical guidance. Gynecol Oncol, 2015. 136(2): p. 178-82. 5. Arbyn, M., et al., Accuracy of human papillomavirus testing on self-collected versus clinician-collected samples: a meta-analysis. Lancet Oncol, 2014. 15(2): p. 172-83. 6. Arbyn, M. and P.E. Castle, Offering Self-Sampling Kits for HPV Testing to Reach Women Who Do Not Attend in the Regular Cervical Cancer Screening Program. Cancer Epidemiol Biomarkers Prev, 2015. 24(5): p. 769-72.

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Benefit for PCR in Human Papillomavirus detection with GenoID PCR technology Overall, there are more than 130 genotypes of HPV. 50 out of these can cause infection in the genital region and 14 are known to be in the high-risk group. For high-quality patient management, it is required to detect all anogenital HPV and especially the high-risk genotypes. A technology that meets these requirements has to be both highly sensitive and specific allowing precise discrimination of HPV types. Only one technology provides this outstanding set of features: PCR, which stands for polymerase chain reaction. DNA based PCR methods are used widely to detect HPV infections. Besides being the most sensitive diagnostic methods, these supply important typing information for counseling, selection of therapy, follow-up and vaccine monitoring. On the basis of this technology GenoID manufactures the Full Spectrum HPV detection kit. The kit is the result of years of research and development and is based on the assay used in GenoID’s clinical laboratory where tens of thousands of HPV tests have been performed. Precise discrimination of the genotypes relies on a unique HPV DNA signature discovered by GenoID’s researchers. Researchers at GenoID have developed a system that features a balanced sensitivity of amplification and an automated, high throughput typing method (hybridization) using the virus L1 gene sequence that is suitable for consensus amplification of HPV types and it is also possible to design both separate general and type-specific probes to the same amplicon. The system allows simultaneous detection and typing of 48 HPV types. The Full Spectrum HPV test service is designed to detect the genital HPV genotypes in the following constitution:

PCR based Human papillomavirus (HPV) group detection & genotyping Genotyping of all HR-HPV types: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68 Genotyping of LR-HPV types: 6, 11 Group detection of LR-HPV types: 42, 43, 44 Group detection of non-classified (NA)-HPV types: 2, 3, 7, 10, 13, 26, 27, 28, 29, 30, 34, 40, 53, 54, 57, 61, 67, 70, 72, 73, 74, 81, 82, 83, 84, 85, 89, 90, 91

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The confidence real-time HPV test Technology The CONFIDENCE™ HPV test (NEUMANN Diagnostics Ltd. CE) is a qualitative in vitro test for the detection of Human Papillomavirus (HPV) in patient specimens. It is a TaqMan-based L1 region specific multiplex real-time PCR assay for viral DNA detection (patent pending). It detects HPV16 and HPV18 separately and other high risk types (HPV31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68) in group with quality controlled high-throughput and highly automated protocols running on Tecan EVO® liquid handling platform using contamination safe workflow. After the 96-channel automated PCR setup, 5-plex quantitative real-time PCR is performed on the QuantStudio™ 6 Flex platform in 384-well plate format. The whole procedure is standardized, and the contamination risk is monitored with a DNA preparation no template control (NTC) and PCR NTC where both results in the PCR plate should be negative for a valid PCR plate run. The quality of the sample DNA is guaranteed by the amplification of the genomic target control and internal control (artificial DNA added to the sample during DNA preparation). The PCR procedures are monitored by PCR positive control.

Validated sample types: 1. Cervical cells collected by the Cervex Brush® Combi (Rovers®) in PreservCyt®Solution (ThinPrep®, Hologic Corp) 2. Self-collected vaginal sample taken with the Qvintip® device (Aprovix).

The clinical performance of the CONFIDENCE™ HPV test was assessed in the prospective TRACE (Triage and Risk Assessment of Cervical Precancer by Epigenetic Biomarker) trial at 4 clinical sites including outpatient and colposcopy clinics in Hungary. The Roche cobas HPV test (Roche Molecular System, CE) was used as a clinically validated reference test with 3,270 ThinPrep® cervical samples of women above 25 years of age. The study endpoint for the analysis was histologically confirmed high-grade CIN, i.e., CIN2 or worse (CIN2+) and CIN3 or worse (CIN3+). The output and comparison of different HPV tests were evaluated by performance and bias adjusted kappa value and positive agreement. The results showed 70.4% agreement in the hrHPV positive cases, which is a very good agreement considering the recent findings in this field [1]. The performance and bias adjusted kappa value [2] of the results was 0.84 (95% CI: 0.82-0.86). The CONFIDENCE™ HPV test for the Qvintip® self-collected vaginal sample was clinically validated by comparing the Confidence HPV test results of the clinician collected ThinPrep® cervical sample with the Qvintip® self-collected vaginal sample. In the TRACE trial 335 women were asked to self-collect a vaginal sample by using the Qvintip® self-sampling kit before the clinician obtained their cervical specimen in ThinPrep®. The results of the two sample types showed similar agreement in the hrHPV positive cases compared to the recent findings in a comparison study of the Qvintip® device [3]. The Confidence HPV test is fully integrated into GenoID‘s laboratory information system. The testing system is able to process up to 600 samples per day.

Intended uses: • First-line primary screening test to identify women at increased risk for the development of cervical cancer or presence of high-grade disease. • To assess the presence or absence of hrHPV types • To assess the presence or absence of hrHPV genotypes 16 and 18.

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References 1. Rebolj, M., et al., Disagreement between human papillomavirus assays: an unexpected challenge for the choice of an assay in primary cervical screening. PLoS One, 2014. 9(1): p. e86835. 2. Xue, X., et al., A new method to address verification bias in studies of clinical screening tests: cervical cancer screening assays as an example. J Clin Epidemiol, 2014. 67(3): p. 343-53. 3. Jentschke, M., et al., Comparative evaluation of two vaginal self-sampling devices for the detection of human papillomavirus infections. J Clin Virol, 2015.

Sexually Transmitted Infections (STI) panel from a single sample STI is short for Sexually Transmitted Infections, and is an umbrella term for various infections caused by different pathogenic microorganisms, which are transmitted from person to person via sexual intercourse.

Sexually transmitted diseases may be caused by various pathogens. These include bacteria, viruses, fungi and other microbae. Most of these infections can be completely cured. Several STIs caused by viruses still pose a problem for treatment, these illnesses can only be treated but not cured. The STI tests can be requested as single assays or combined in different groups, for example 8+1 (including HPV test). Due to the common technological platform developed by GenoID, one sampling is sufficient (cervix, urine, semen etc.) to perform 9 tests. The Treponema pallidum test can only be performed from genital ulcer scraping sample.

A range of illnesses may be transmitted with sexual intercourse, all with different signs and symptoms. Their incubation period varies from several hours to months, and often do not have any visible signs or symptoms. If the infection is mostly limited to the genitals, the symptoms may include abnormal urethral or vaginal discharge, sores, blisters, various growths, pain or itching. In other cases there are no noticeable symptoms for quite some time, despite the fact that the individual has been infected and is able to pass it on. In these cases, STI testing may be employed to spot the infection before the onset of effects and symptoms. An accurate diagnosis is vital for proper treatment. Post-treatment tests are just as important to determine if the patient is completely healthy. The diminishment or disappearance of symptoms does not necessarily mean that the infection has been eliminated. Left untreated, these diseases lead to serious complications. Pathogens can attack the internal genitalia in both men and women, causing infertility. In the case of pregnancy, the parents’ STI poses a risk for the unborn child, because during pregnancy, birth or breastfeeding the mother may infect the embryo or child. There are also STIs which cause secondary inflammations of the joints and eyes, as well as skin diseases, which are even harder to treat.

• Chlamydia trachomatis (CT) • Neisseria gonorrhoeae (NG) • Mycoplasma genitalium (MG) • Ureaplasma urealyticum/parvum (UU/UP) • Trichomonas vaginalis (TV) • Gardnerella vaginalis (GV) • Atopobium vaginae (AV) • Herpes simplex virus1/2 (HSV1/2) • Treponema pallidum (TP)

9 tests from 1 sample presents several advantages to both the practitioner and the patient: • Provides complete “women’s and men’s health” results • Comprehensive and detailed picture about the clinical status of the patient in case of vaginal discharge, cervicitis, urethritis, prostatitis • Multiple sampling has disadvantages as it is painful and expensive, requires multiple validated detection systems with all the extra costs involved.

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Vaginal discharge/cervicitis/urethritis/prostatitis vaginitis vaginosis

STI and HPV screening

Complete panel

First screening

CT

+

+

+

NG

+

+

+

MG

+

+

+

UU/UP

+

TV

+

+

+

GV

+

+

+

AV

+

+

+

HSV1/2

+

Cervical cancer screening

+

+

+ +

TP HPV

Genital ulceration

+

+

+

+

+

The HPV test may be requested together with all panels, except for the genital ulcer panel.

Tests from blood sample HIV/HCV/HBV PCR screening test (cobas s201 MPX test 2.0, Roche, CE) is intended for the screening of blood and other organ donations, and is not intended for diagnosis. We accept samples with these indications only.

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A short for the collecton, storage and transport of samples for testing at GenoID laboratory How to collect and store STI samples GenoID’s automated system is able to handle many kinds of male and female sample types, providing results to a wide range of clinicians such as gynaecologists, dermatologists, urologists and researchers in 24-72 hours. Several transport media to preserve cells for pathogen detection have been validated on GenoID’s system [Table]. Various sample types are handled in different ways to assure that the samples arrive in their best conditions for diagnostics.

Sample tube or swab name

Manufacturer

Storage

Stability

Transport

Valid for

CyMol

Copan

2-30°C

3 weeks

ground only, flammable

STD+HPV

Easyfix

Labonord

2-30°C

3 weeks

ground only, flammable

STD+HPV

LiquiPrep

LGM International Inc.

2-30°C

1 week

ground only, flammable

STD+HPV

SurePath

Becton, Dickinson and Company

2-30°C

3 weeks

ground only, flammable

STD+HPV

PreservCyt

Hologic Corp.

2-30°C

3 weeks

ground only, flammable

STD+HPV

cobas® PCR Cell Collection Media

Roche Molecular Systems Inc.

15-30°C

6 weeks

ground only, flammable

STD+HPV

cobas® PCR Female Swab Sample Kit

Roche Molecular Systems Inc.

2-30°C

3 weeks

air or ground transport

STD+HPV

cobas® PCR Urine Sample Kit

Roche Molecular Systems Inc.

2-30°C

3 weeks

air or ground transport

STD

buccal swab (brush type only)

any CE marked

2-30°C

3 weeks

air or ground transport

all genetic tests

forensic dry swab

Copan Flocked Swab 4N6, snap off, 2 ml eppendorf tube with evaporation-cap, peelpack, sterile; Code: M10181

2-30°C

3 weeks

air or ground transport

all genetic tests

cervical swab or brush

any CE marked for cervical sampling

NA

NA

NA

STD+HPV

urethral swab or brush

any CE marked for urethral sampling

NA

NA

NA

STD+HPV

urine sample collection tube

any sterile CE marked

NA

NA

NA

STD

Qvintip®

Aprovix

2-30°C

1 week

air or ground transport

HPV

Table: Summary of sample collection, storage and transport instructions for GenoID tests

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Sample types validated for STD and HPV tests • Cervical sample: The cervical smear taking protocol should be followed. The swab or brush sample should be transferred into any of the following transport tubes: Cymol, Easyfix, Liquiprep, Surepath, PreservCyt, cobas® PCR Cell Collection Media, cobas® PCR Female Swab Sample Kit. An aliquot from the original tubes with a minimum volume of 3 ml is needed. • Urine: first catch urine, minimum 8 ml collected in a sterile urine cup. • Native urine: may be stored at room temperature for 24h, and for 6 months when frozen. Store frozen until shipping. • Stabilized urine: Urine sample may be stabilized with cobas® PCR Urine Sample Kit and transported at room temperature. Once the specimen has been stabilized it, may be stored at 2-30ºC for up to 90 days. • Semen: minimum 0.5-1 ml collected in a sterile urine cup. Store frozen until shipping. • Urethral swab or brush sample: should be transferred into any of the transport tubes listed at cervix • Vaginal swab or brush sample: should be transferred into any of the transport tubes listed at cervix • Pharyngeal swab or brush sample (for STD only): should be transferred into any of the transport tubes listed at cervix • Anal swab or brush sample: should be transferred into any of the transport tubes listed at cervix • Scraping, lesion scraping: swab or brush sample should be transferred into any of the transport tubes listed at cervix • Condyloma: swab sample or biopsies should be transferred into any of the transport tubes listed at cervix. Please mark “Biopsy” on the tube as well. • Qvintip®: vaginal self sampling, see the Qvintip® sample collection guide Upon request we validate other sample transport tubes and sample types.

Please contact us for further details.

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Examples for sample collection devices and transport systems • ThinPrep (PreservCyt) (Hologic, CE) liquid based cytology (LBC) transport tube • broom device for cervical sample collection • brush device for cervical sample collection • cervical sampling devices

More detailed instructions on sample collection and handling can be found at these links: Thinprep: http://www.thinprep.com/hcp/specimen_collection/broom_like_device.html cobas® PCR Female Swab Sample Kit, cobas® PCR Urine Sample Kit: http://www.roche-diagnostics.ch/content/dam/corporate/roche-dia_ch/documents/broschueren/molecular_diagnostics/hpv _microbiology/PCR_SampleKitBroch_A4_FINAL.pdf

How to request tests For our partner’s convenience we accept any kind of electronic or paper request forms that contain the necessary information. Alternatively, you can download our test request form from our website: http://www.genoid.net/services/test-request-form

How to send samples Many international courier companies are available to deliver packages safely and fast to GenoID. It takes 24 hours to receive samples from Europe and 2-3 days from the rest of the world. There are special regulations by IATA that should be followed when sending biological samples by air that can easily be followed. Our Referral Manager assists you from the first steps.

How to receive results Turnaround times for tests are 48-72 hours (on working days) from receipt in the laboratory. Different report formats are available to best serve our partners’ needs. You may choose to have the reports in the following way: • PDF/Excel form • GenoID’s Online Result System • Fax

SYNLAB Budapest Diagnostic Center GenoID Molecular Diagnostic Laboratory Szanatórium u. 19. H-1121 Budapest, Hungary Tel.: 00 36 1 465 01 24 Fax: 00 36 1 465 01 27 [email protected]

www.genoid.net SYNLAB Hungary Kft. Bajcsy-Zsilinszky út 53. 1st fl. H-1065 Budapest, Hungary

www.synlab.hu

© SYNLAB Hungary Kft. The company excludes all liability for printing errors, mistakes or any inaccuracy of this text and reserves the right to change, update or modify it. All texts, contents and images are under copyright. You may not, except with expressed written permission of intellectual property rights holders, use, distribute or exploit them commercially. As of 06/2016

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