I N S T A N D e. V. Gesellschaft zur Förderung der Qualitätssicherung in medizinischen Laboratorien e. V. (vormals Hämometerprüfstelle)

Transcription

1 I N S T A N D e. V. Gesellschaft zur Förderung der Qualitätssicherung in medizinischen Laboratorien e. V. (vormals Hämometerprüfstelle) WHO Collaborating Centre for Quality Assurance and Standardization in Laboratory Medicine in cooperation with Deutsche Vereinigung zur Bekämpfung der Viruskrankheiten (DVV) Gesellschaft für Virologie (GfV) Deutsche Gesellschaft für Hygiene und Mikrobiologie (DGHM) INSTAND-Geschäftsstelle Ubierstr Düsseldorf Telefon: +49 (0) Fax: +49 (0) instand@instand-ev.de Internet: EQAS Adviser: Assistant EQAS Adviser: Prof. Dr. Heinz Zeichhardt Dr. Hans-Peter Grunert CharitéCentrum für diagnostische und präventive Labormedizin Institut für Biotechnologische Diagnostik in der GBD Institut für Virologie, Campus Benjamin Franklin Potsdamer Chaussee 80, Berlin Hindenburgdamm 27, Berlin Tel.: +49-(0) ; Fax: +49-(0) Tel.: +49-(0) /23; Fax: +49-(0) HPGrunert@gmx.de Heinz.Zeichhardt@charite.de February 2012 Final Report External Quality Assessment Scheme (EQAS) - November/December 2011 Virus Genome Detection - Human Papilloma Viruses (373) Differentiation of "High Risk" and "Low Risk" Types and Typing (PCR/NAT HPV) INSTAND-Target Value Laboratories: Uniklinik Köln, Institut für Virologie, Nationales Referenzzentrum für Papillom- und Polyomaviren: Prof. Dr. H. Pfister, Prof. Dr. U. Wieland, Dr. R. Kaiser Charité - Universitätsmedizin Berlin, Campus Mitte, Institut für Medizinische Virologie, Nationales Konsiliarlaboratorium für Hantaviren: Prof. Dr. D. H. Krüger, PD Dr. J. Hofmann Klinikum der Johann Wolfgang Goethe-Universität, Institut für Medizinische Virologie, Frankfurt/Main: Prof. Dr. H. W. Doerr, Prof. Dr. H. Rabenau, PD Dr. A. Berger, Dr. R. Allwinn Labor Enders, Institut für Virologie, Infektiologie und Epidemiologie, Stuttgart: Prof. Dr. Gisela Enders & Partner Universitätsklinikum des Saarlandes, Institut für Virologie, Homburg/Saar; Prof. Dr. S. Smola, Prof. Dr. N. Müller-Lantzsch 373 Human Papilloma Viruses Genome November December 2011 Letter a.doc 1

2 Dear colleagues, Attached please find the final report for the EQAS "Virus Genome Detection - Human Papilloma Viruses" together with the participation documents - certificate of successful participation, - statement of participation, - statement of individual. As you have already been informed, several alterations have become effective with the new Guideline of the German Medical Association (RiliBÄK). In this final evaluation we may again inform you about the following topics: 1 New RiliBÄK (Guideline of the German Medical Association) and validity period of certificate of the INSTAND EQA schemes 1.1 New RiliBÄK (Guideline of the German Medical Association) The new Guideline of the German Medical Association (Bundesärztekammer / RiliBÄK = Richtlinie der Bundesärztekammer zur Qualitätssicherung laboratoriumsmedizinischer Untersuchungen) specifies the basic principles of internal and external quality assurance of all laboratory medical analyses including those in virus diagnostics (please see: Deutsches Ärzteblatt, Jg. 108, Heft 30, , Seite A ). All qualitative and quantitative analyses in virus diagnostics are subject to internal quality assurance. In addition all analyses in virus diagnostics which are specified for external quality assurance (external quality assessment/eqa schemes) in the tables of the different RiliBÄK sections are subject to mandatory participation in the corresponding EQA scheme. The quality assurance of tests for the detection of virus specific antibodies was assigned to section B 2 of the RiliBÄK"qualitative determinations in laboratory medicine = Qualitative laboratoriumsmedizinische Untersuchungen" (effective since with a transition period of two years). Reporting a result of a test for virus specific antibody detection primarily qualitatively as "positive", "" or "borderline/indeterminate" implies that the quality assurance of the corresponding test has to be performed according to RiliBÄK section B 2 (qualitative determinations in laboratory medicine = Qualitative laboratoriumsmedizinische Untersuchungen). In this case quantitative statements on antibody concentrations* (i.e. as units/volume) can additionally be reported in brackets for orientation. Please note that primary reporting of antibody concentration alone or in front of the qualitative result directly implies that the quality assurance of the corresponding test has to be performed according to RiliBÄK section B 1 ("quantitative determinations in laboratory medicine = Quantitative laboratoriumsmedizinische Untersuchungen"). Table B 2-2 of section B 2 (external quality assurance = Externe Qualitätssicherung/Ringversuche) comprises tests for the detection of antibodies for which participation in the corresponding EQA schemes is mandatory. This concerns testing of antibodies against HIV, antibodies against hepatitis A virus, antibodies against hepatitis B virus (anti-hbc, anti-hbe, anti-hbs), antibodies against hepatitis C virus, antibodies against rubella virus. RiliBÄK defines the validity period of a certificate for each of the above mentioned EQA schemes stated in Table B 2-2 (Externe Qualitätssicherung/Ringversuche) as twice as long as the frequency for mandatory participation. All INSTAND EQA schemes in virus diagnostics which are not subject of table B 2-2 will be organized by INSTAND in compliance to RiliBÄK. The quality assurance of all tests for the direct virus detection (by microscopy and cell culture as well as molecular biological and immunological procedures) will be defined in RiliBÄK section B 3 (direct detection and characterization of infectious diseases pathogens = Direkter Nachweis und Charakterisierung von Infektionserregern) which is in progress. The validity period of the certificates of the virological INSTAND EQA schemes for tests stated in section B 3 will follow the already implemented RiliBÄK section B 2 (qualitative determinations in laboratory medicine = Qualitative laboratoriumsmedizinische Untersuchungen). * Please note that determination of titers is considered as qualitative analysis (see section A of RiliBÄK "General requirements on quality assurance of laboratory medical analyses" = "Grundlegende Anforderungen an die Qualitätssicherung laboratoriumsmedizinischer Untersuchungen"). 373 Human Papilloma Viruses Genome November December 2011 Letter a.doc 2

3 1.2 Validity period of certificate of the INSTAND EQA schemes Based on the above described principles in section 1.1 it has been decided that the validity period of the certificates of successful participation of all INSTAND EQA schemes in virus diagnostics will be one year which has been started with the EQA schemes in September This concerns the EQA schemes in virus diagnostics performed with a frequency of four times per year (Table 1) as well as those schemes performed twice a year (Table 2). The validity period of certificate will be two years for EQA schemes performed once a year. The validity period of the certificate of the "Special EQA program in accordance with the RKI-entero surveillance program - Virus Detection - Enterovirus-PCR / cultivation and typing" (program 374) will be agreed with the National Reference Center for Poliomyelitis and Enteroviruses, Regional Reference Laboratory of WHO/EURO for Poliomyelitis, Robert Koch-Institute, Berlin. 2 Mailing of documents of a defined EQA scheme term Certificate of successful participation, statement of participation and statement of individual The certificates of successful participation, statements of participation and statements of individual of the INSTAND EQA schemes in virus diagnostics performed with a frequency of four times a year (Table 1) will be mailed as printouts together with the pre-evaluation. This concerns the following programs: Table 1 Mailing of documents together with the pre-evaluation EQA schemes in virus diagnostics performed with a frequency of four times a year Virus immunology: 335 HIV-1/HIV HIV-1 p24 Ag 343 Hepatitis A virus 344 Hepatitis B virus (Prog. I) 345 Hepatitis B virus (Prog. II) 346 Hepatitis C virus 351 Cytomegalovirus Virus genome detection: 360 HIV-1 (RNA) 361 Hepatitis B virus 362 Hepatitis C virus 365 Cytomegalovirus 367 Parvovirus B Hepatitis A virus The final evaluations** of the EQA programs stated in Table 1 will be mailed independently. The procedure of mailing of the documents (certificates of successful participation, statements of participation and statements of individual ) of the INSTAND EQA schemes in virus diagnostics performed twice a year or with lower frequency remains unchanged for a defined EQA scheme term (Table 2). As recently practiced, the documents will be mailed together with the printouts of the final evaluations**. & Table 2 Mailing of documents together with the final evaluations** EQA schemes in virus diagnostics performed twice a year or with lower frequency Virus immunology: 341 Rubella virus 342 Parvovirus B Hepatitis D virus 348 Hepatitis E virus 350 Dengue viruses (Ab/NS1-Ag) 352 Epstein Barr virus 353 Varicella zoster virus 354 Herpes simplex viruses 355 Hantaviruses 356 Mumps virus 357 Measles virus 358 TBE (FSME) virus 359 Respiratory syncytial virus (Ag/genome) Virus genome detection: 363 Herpes simplex virus 1/2 366 Varicella zoster Virus 370 Influenza viruses (genome/ag) & 371 Adenoviruses 372 Enteroviruses 373 Human Papilloma viruses 374 RKI-Entero-Surveillance (every two years) 375 HCV genotyping (once a year) 376 Epstein Barr virus The two terms of the EQA scheme "Influenza viruses" (370) have been appointed to the winter season, i.e. November and March of the following year. The procedure of mailing of documents (certificates of successful participation, statements of participation and statements of individual ) and final evaluation of the influenza EQA scheme of the November term follows the specifications as described for the EQA schemes in Table 1. ** As already recently practiced, the printouts of the final evaluations will only be mailed to you in case you have already registered for this procedure at INSTAND. Please note that all final evaluations of the EQA schemes in virus diagnostics since 2006 are available as PDF file on the INSTAND-homepage (see section 3). 373 Human Papilloma Viruses Genome November December 2011 Letter a.doc 3

4 3 Release of final evaluations of EQA schemes on the INSTAND homepage Each final evaluation of a defined EQA scheme will be released on the INSTAND homepage immediately after completion. The final evaluations as well as the pre-evaluation of this EQA scheme are available as PDF file on the INSTAND-homepage under EQAS / Reports / Year and Category (Virus genome detection) in English language ( ) and in German language ( ). 4 Test categories, statement of and evaluation criteria for this EQA scheme As already recently practiced, the certificate of successful participation of a defined EQA scheme in virus diagnostics is assigned to a defined test category. This EQA scheme comprises the following test categories (Table 3): Table 3 Test categories and statement of Statement of Test categories (the following statements of were requested) (20) Differentiation of "High Risk" and "Low Risk" s of HPV (human papilloma viruses) - Differenzierung von HPV (Humane Papillomviren) in "High Risk"- und "Low Risk"-Typen (25) Typing of "High Risk" HPV and detection of "Low Risk" HPV (without typing) - Typisierung von "High Risk"-HPV und Nachweis von "Low Risk"-HPV (ohne Typisierung) (30) Typing of "High Risk" HPV and "Low Risk" HPV - Typisierung von "High Risk"-HPV und "Low Risk"-HPV (35) Typing of "High Risk" HPV (without detection of "Low Risk" HPV) - Typisierung von "High Risk"-HPV (ohne Bestimmung von "Low Risk"-HPV) Comment 1: These tests exclusively detect and "High Risk" HPV and cannot detect "Low Risk" HPV. This means that a test result obtained by these tests can only be interpreted as " for 'High Risk' HPV". Results representing " for 'High Risk' HPV" will be stated as "" in your statement of individual. (60) Detection of "High Risk" HPV (without detection of "Low Risk" HPV) - Bestimmung von "High Risk"-HPV (ohne Bestimmung von "Low Risk"-HPV) Comment 2: These tests exclusively detect "High Risk" HPV (without specification of the HPV ) and cannot detect "Low Risk" HPV. This means that a test result obtained by these tests can only be interpreted as " for 'High Risk' HPV". Results representing " for 'High Risk' HPV" will be stated as "" in your statement of individual. Comment 3: Tests for the detection of "High Risk HPV E6/E7 mrna were assigned to test category 60 because this test system only detects E6/E7 mrna of a distinct number of "High Risk" HPV s. Statement of - "High Risk", - "Low Risk", - " " or - "indeterminate" Detection of - "High Risk" HPV reporting of the detected HPV : e.g. ""; - "Low Risk" HPV reporting of "Low Risk"; - samples reporting of " " Detection of - "High Risk" HPV reporting of the detected HPV : e.g. ""; - "Low Risk" HPV reporting of the detected HPV : e.g. "HPV 6"; - samples reporting of " " Detection of - "High Risk" HPV reporting of the detected HPV : e.g. ""; - " for 'High Risk' HPV" (i.e. samples, which are positive for "Low Risk"-HPV or samples, which are for HPV) reporting of " for 'High Risk' HPV" (see comment 1) Detection of - "High Risk" HPV reporting of "High Risk" - " for 'High Risk' HPV" (i.e. samples, which are positive for "Low Risk"-HPV or samples, which are for HPV) reporting of " for 'High Risk' HPV" (see comments 2 and 3) 373 Human Papilloma Viruses Genome November December 2011 Letter a.doc 4

5 A result was not considered for evaluation in case you had specified that this result should only be taken as additional information and ignored as valid result. The test categories 20, 25, 30, 35 and 60, respectively, are individually evaluated in the certificate of successful participation and listed in all participation and evaluation documents. The evaluation criteria for the of EQA schemes for the detection of virus specific antibodies follow the new Guideline of the German Medical Association, RiliBÄK section E 2 (specific requirements on EQA schemes for qualitative laboratory medical analyses = Spezielle Anforderungen an Ringversuche bei qualitativen laboratoriumsmedizinischen Untersuchungen). For receiving a certificate of successful participation it is required that you analyzed all samples of the sample set ly with the same method in the corresponding test categories (100 according to the target values). A corresponding proceeding is applied for tests for virus antigen and genome detection. Example - Program "Virus Genome Detection - Human Papilloma Viruses" (373): All 5 samples of the sample set have to be tested ly with the same method in test category 20 "Differentiation of "High Risk" and "Low Risk" s of HPV (human papilloma viruses) - Differenzierung von HPV (Humane Papillomviren) in "High Risk"- und "Low Risk"-Typen ". The same applies for test categories 25, 30, 35 and 60 of this EQA scheme. 5 Sample properties, target values, and success rates Please see Table 4 for details on the properties of the samples of this EQA scheme. Sample No. Table 4: Sample properties and target values Qualitative Type Dilution Origin for HPV Lysate of MRC-5 cells (human lung fibroblasts), no HPV genome detectable * High Risk 1:5* Lysate of SiHa cells, containing sequences of # without evaluation # expected result: Low Risk without evaluation # expected result: HPV 42, 61 and s 1:50 Biopsy material of a condyloma High Risk HPV 18 1:30 Lysate of HeLa cells, containing sequences of HPV * High Risk 1:10* Lysate of SiHa cells, containing sequences of. * The positive samples and represented different dilution steps of a dilution series of the same lysate of SiHa cells. # The reported for sample were inconsistent and therefore not evaluated (without disadvantage for your certificate of successful participation). The are summarized and differentiated for each sample according to test category 20, 25, 30, 35 and 60, respectively, in Table 5. Detailed for the detection and differentiation between "High Risk" and "Low Risk" HPVs as well as for typing are shown in the attached tables. We gratefully acknowledge the good cooperation with Prof. Dr. H. Pfister, Prof. Dr. U. Wieland and Dr. R. Kaiser (Nationales Referenzzentrum für Papillom- und Polyomaviren, Institut für Virologie, Universität Köln) as well as Prof. Dr. H.-P. Berlien and Dr. U. Müller (Evangelische Elisabeth Klinik, Berlin). We additionally thank Dr. M. Enders and Dr. G. Schalasta (Labor Prof. Gisela Enders und Partner, Stuttgart) for the studies on suitability of the EQA samples for the detection of "High Risk" HPV E6/E7 mrna by a TMA test system. Thank you very much for your kind cooperation. Prof. Dr. H. Zeichhardt 373 Human Papilloma Viruses Genome November December 2011 Letter a.doc 5

6 Table 5: Summary of sample properties, target values, and success rates Sample Sample * Sample # Sample Sample * Properties HPV HPV HPV HPV HPV positive positive # positive positive High Risk or expected result: High Risk # Low Risk Low Risk High Risk High Risk Type 16 expected result: # s 42, 61 and s Material lysate of MRC-5 cells lysate of SiHa cells biopsy material lysate of HeLa cells lysate of SiHa cells Dilution : 5* 1 : 50 1 : 30 1 : 10* considered as "" result (target value - test category 20: differentiation of "High Risk" and "Low Risk" s of HPV) High Risk without evaluation # High Risk High Risk (Low Risk positive : (86/87) (86/87) /87) (85/87) (86/87) considered as "" result (target value - test category 25: typing of "High Risk" HPV and detection of "Low Risk" HPV (without typing)) / not done without evaluation # HPV (Low Risk positive : (9/9) (9/9) /9) (9/9) (9/9) considered as (target value - test category 30: typing of "High Risk" HPV and "Low Risk" HPV) / not done without evaluation # HPV (62/62) 100 (62/62) (HPV 42, 61 positive : /62) Success rates for all 5 samples of the sample set & 373 Human Papilloma Viruses Genome November December 2011 Letter a.doc (62/62) 100 (62/62) considered as "" result (target value - test category 35: typing of "High Risk" HPV (without detection of "Low Risk" HPV)) for High Risk-HPV without evaluation # HPV (15/15) 100 (15/15) ( for High Risk HPV : /15) 100 (15/15) 100 (15/15) considered as "" (target value - test category 60: detection of "High Risk" HPV (without detection of "Low Risk" HPV)) for High Risk-HPV High Risk without evaluation # High Risk High Risk 100 (22/22) 100 (22/22) ( for High Risk HPV : /22) * The positive samples and represented different dilution steps of a dilution series of a lysate of SiHa cells. # The reported for sample were inconsistent and therefore not evaluated (without disadvantage for your certificate of successful participation). & The success rates for all 5 samples of the corresponding sample set of test categories 20, 25, 30, 35 and 60, respectively, refer to the number of participating laboratories. Laboratories having reported obtained by several methods are recorded only once in the corresponding test category. 100 (22/22) 100 (22/22) 96.5 &# (83/86) 100 &# (9/9) 100 &# (59/59) 100 &# (15/15) 100 &# (22/22)

7 INSTAND e. V., Gesellschaft zur Förderung der Qualitätssicherung in medizinischen Laboratorien e. V. (vormals Hämometerprüfstelle) - in cooperation with Deutsche Vereinigung zur Bekämpfung der Viruskrankheiten e. V. (DVV) Gesellschaft für Virologie e. V. (GfV) Deutsche Gesellschaft für Hygiene und Mikrobiologie e. V. (DGHM) EQAS Virology November/December 2011 Virus genome detection PCR- / NAT-HPV (Human Papilloma Viruses) (373) Results for sample Differentiation of "High Risk" and "Low Risk" s of HPV Correct result: test in High Risk Low Risk indeterminate of High Risk Low Risk indeterminate of hybridization (70) Gen ID (AID) - HPV-screening Innogenetics - INNO-LiPA HPV Genotyp. Extra Gen ID (AID) - HPV-typing Greiner - PapilloCheck HPV Screening Roche - Linear Array HPV Genotyping Test in house Chipron - HPV Type nested PCR / hybridization (1270) in house Abbott - RealTime High Risk HPV hybridization / hybridization (3070) Qiagen - digene HC2 HPV DNA Test sequencing (72) in house nested PCR / sequencing (1272) in house test (99) PCR / test (1199) in house nested PCR / test (1299) in house Multiplex PCR / test (6199) manufacturer Human Papilloma Viruses Genome November December 2011 Table b.doc 1

8 Results for sample Typing of "High Risk" HPV and detection of "Low Risk" HPV (without typing) Correct result: test in hybridization (70) Gen ID (AID) - HPV-screening Gen ID (AID) - HPV-typing Results for sample Typing of "High Risk" HPV and "Low Risk" HPV Correct result: test in hybridization (70) Innogenetics - INNO-LiPA HPV Genotyp. Extra Greiner - PapilloCheck HPV Screening Roche - Linear Array HPV Genotyping Test Chipron - HPV Type in house Gen ID (AID) - HPV-typing nested PCR / hybridization (1270) in house Abbott - RealTime High Risk HPV PCR / hybridization (9870) Sacace - HPV High Risk Typing Human Papilloma Viruses Genome November December 2011 Table b.doc 2

9 Results for sample Typing of "High Risk" HPV and "Low Risk" HPV (continued) sequencing (72) in house nested PCR / sequencing (1272) in house test (99) PCR / test (1199) in house Multiplex PCR / test (6199) manufacturer Results for sample Typing of "High Risk" HPV (without detection of "Low Risk" HPV) (Tests for exclusive detection and typing of "High Risk"-HPV )* Correct result: for "High Risk"-HPV test in * * hybridization (70) Abbott - RealTime High Risk HPV sequencing (72) in house test (99) PCR / test (1199) AmpliSens - HPV-HCR-geno EPh TaqMan / test (1399) Roche - Cobas 4800 HPV Test manufacturer * These tests exclusively detect and "High Risk"-HPV and cannot detect "Low Risk"-HPV. This means that a test result obtained by these tests can only be interpreted as " for High Risk-HPV". Results representing " for High Risk-HPV" will be stated as "" in your statement of individual. 373 Human Papilloma Viruses Genome November December 2011 Table b.doc 3

10 Results for sample Detection of "High Risk" HPV (without detection of "Low Risk" HPV) (Tests which exclusively detect "High Risk"-HPV)* Correct result: for "High Risk"-HPV test in High Risk Low Risk indeterminate of * High Risk Low Risk indeterminate of * hybridization (70) Roche - Amplicor HPV Test Abbott - RealTime High Risk HPV hybridization / hybridization (3070) Qiagen - digene HC2 HPV DNA Test sequencing (72) in house test (99) PCR / test (1199) manufacturer TaqMan / test (1399) manufacturer TMA / test (6099) Gen-Probe - Aptima HPV Assay # * These tests exclusively detect "High Risk"-HPV (without specification of the HPV ) and cannot detect "Low Risk"-HPV. This means that a test result obtained by these tests can only be interpreted as " for High Risk-HPV". Results representing " for High Risk-HPV" will be stated as "" in your statement of individual. # This test system detects E6/E7 mrna of a distinct number of "High Risk" HPV s (see Table 3, Comment 3). 373 Human Papilloma Viruses Genome November December 2011 Table b.doc 4

11 Results for sample Differentiation of "High Risk" and "Low Risk" s of HPV Correct result: High Risk test in High Risk Low Risk indeterminate of High Risk Low Risk indeterminate of hybridization (70) Gen ID (AID) - HPV-screening Innogenetics - INNO-LiPA HPV Genotyp. Extra Gen ID (AID) - HPV-typing Greiner - PapilloCheck HPV Screening Roche - Linear Array HPV Genotyping Test in house Chipron - HPV Type nested PCR / hybridization (1270) in house Abbott - RealTime High Risk HPV hybridization / hybridization (3070) Qiagen - digene HC2 HPV DNA Test sequencing (72) in house nested PCR / sequencing (1272) in house test (99) PCR / test (1199) in house nested PCR / test (1299) in house Multiplex PCR / test (6199) manufacturer Human Papilloma Viruses Genome November December 2011 Table b.doc 5

12 Results for sample Typing of "High Risk" HPV and detection of "Low Risk" HPV (without typing) Correct result: test in hybridization (70) Gen ID (AID) - HPV-screening Gen ID (AID) - HPV-typing Results for sample Typing of "High Risk" HPV and "Low Risk" HPV Correct result: test in hybridization (70) Innogenetics - INNO-LiPA HPV Genotyp. Extra Greiner - PapilloCheck HPV Screening Roche - Linear Array HPV Genotyping Test Chipron - HPV Type in house Gen ID (AID) - HPV-typing nested PCR / hybridization (1270) in house Abbott - RealTime High Risk HPV PCR / hybridization (9870) Sacace - HPV High Risk Typing Human Papilloma Viruses Genome November December 2011 Table b.doc 6

13 Results for sample Typing of "High Risk" HPV and "Low Risk" HPV (continued) sequencing (72) in house nested PCR / sequencing (1272) in house test (99) PCR / test (1199) in house Multiplex PCR / test (6199) manufacturer Results for sample Typing of "High Risk" HPV (without detection of "Low Risk" HPV) (Tests for exclusive detection and typing of "High Risk"-HPV )* Correct result: test in * * hybridization (70) Abbott - RealTime High Risk HPV sequencing (72) in house test (99) PCR / test (1199) AmpliSens - HPV-HCR-geno EPh TaqMan / test (1399) Roche - Cobas 4800 HPV Test manufacturer * These tests exclusively detect and "High Risk"-HPV and cannot detect "Low Risk"-HPV. This means that a test result obtained by these tests can only be interpreted as " for High Risk-HPV". Results representing " for High Risk-HPV" will be stated as "" in your statement of individual. 373 Human Papilloma Viruses Genome November December 2011 Table b.doc 7

14 Results for sample Detection of "High Risk" HPV (without detection of "Low Risk" HPV) (Tests which exclusively detect "High Risk"-HPV)* Correct result: High Risk test in High Risk Low Risk indeterminate of * High Risk Low Risk indeterminate of * hybridization (70) Roche - Amplicor HPV Test Abbott - RealTime High Risk HPV hybridization / hybridization (3070) Qiagen - digene HC2 HPV DNA Test sequencing (72) in house test (99) PCR / test (1199) manufacturer TaqMan / test (1399) manufacturer TMA / test (6099) Gen-Probe - Aptima HPV Assay # * These tests exclusively detect "High Risk"-HPV (without specification of the HPV ) and cannot detect "Low Risk"-HPV. This means that a test result obtained by these tests can only be interpreted as " for High Risk-HPV". Results representing " for High Risk-HPV" will be stated as "" in your statement of individual. # This test system detects E6/E7 mrna of a distinct number of "High Risk" HPV s (see Table 3, Comment 3). 373 Human Papilloma Viruses Genome November December 2011 Table b.doc 8

15 Results for sample Differentiation of "High Risk" and "Low Risk" s of HPV Correct result: without evaluation test in High Risk Low Risk indeterminate of High Risk Low Risk indeterminate of hybridization (70) Gen ID (AID) - HPV-screening Innogenetics - INNO-LiPA HPV Genotyp. Extra Gen ID (AID) - HPV-typing Greiner - PapilloCheck HPV Screening 5 5 Roche - Linear Array HPV Genotyping Test 4 4 in house Chipron - HPV Type nested PCR / hybridization (1270) in house 1 1 hybridization / hybridization (3070) Qiagen - digene HC2 HPV DNA Test PCR / hybridization (9870) altona Diagn. - RealStar HPV LR PCR 4 4 sequencing (72) in house nested PCR / sequencing (1272) in house test (99) PCR / test (1199) in house 1 1 nested PCR / test (1299) in house 1 1 Multiplex PCR / test (6199) manufacturer Human Papilloma Viruses Genome November December 2011 Table b.doc 9

16 Results for sample Typing of "High Risk" HPV and detection of "Low Risk" HPV (without typing) Correct result: without evaluation test in Low Risk Low Risk hybridization (70) Gen ID (AID) - HPV-screening Gen ID (AID) - HPV-typing Results for sample Typing of "High Risk" HPV and "Low Risk" HPV Correct result: without evaluation test in HPV 42 HPV HPV 42 HPV 61 hybridization (70) Innogenetics - INNO-LiPA HPV Genotyp. Extra Greiner - PapilloCheck HPV Screening 8 8 Roche - Linear Array HPV Genotyping Test 15 6 a, b c, d Chipron - HPV Type e 1 1 in house 5 3 f 2 Gen ID (AID) - HPV-typing 4 4 nested PCR / hybridization (1270) in house 1 1 PCR / hybridization (9870) altona Diagn. - RealStar HPV LR PCR 1 1 Sacace - HPV High Risk Typing 1 1 g a Five participants detected HPV 56. b One participant detected HPV s 18, 56 and 45. c One participant detected additionally HPV s 42 and 83. d One participant detected additionally HPV 42. e One participant detected additionally HPV 61. f One participant detected additionally HPV s 16, 45 and 56. g One participant detected HPV Human Papilloma Viruses Genome November December 2011 Table b.doc 10

17 Results for sample Typing of "High Risk" HPV and "Low Risk" HPV (continued) HPV 42 HPV 61 sequencing (72) in house h, i nested PCR / sequencing (1272) in house j, k test (99) PCR / test (1199) in house Multiplex PCR / test (6199) manufacturer 2 1 l 1 h One participant detected HPV 90. i One participant detected HPV 16. j Two participants detected HPV HPV 81. k One participant detected HPV 83. l One participant detected HPV 56. Results for sample Typing of "High Risk" HPV (without detection of "Low Risk" HPV) (Tests for exclusive detection and typing of "High Risk"-HPV )* Correct result: without evaluation test in hybridization (70) Abbott - RealTime High Risk HPV sequencing (72) in house 1 1 test (99) PCR / test (1199) AmpliSens - HPV-HCR-geno EPh 1 1 TaqMan / test (1399) Roche - Cobas 4800 HPV Test 2 2 manufacturer 1 1 * * * These tests exclusively detect and "High Risk"-HPV and cannot detect "Low Risk"-HPV. This means that a test result obtained by these tests can only be interpreted as " for High Risk-HPV". Results representing " for High Risk-HPV" will be stated as "" in your statement of individual. 373 Human Papilloma Viruses Genome November December 2011 Table b.doc 11

18 Results for sample Detection of "High Risk" HPV (without detection of "Low Risk" HPV) (Tests which exclusively detect "High Risk"-HPV)* Correct result: without evaluation test in High Risk Low Risk indeterminate of * High Risk Low Risk indeterminate of * hybridization (70) Roche - Amplicor HPV Test 2 2 Abbott - RealTime High Risk HPV hybridization / hybridization (3070) Qiagen - digene HC2 HPV DNA Test 5 5 sequencing (72) in house 1 1 test (99) PCR / test (1199) manufacturer 2 2 TaqMan / test (1399) manufacturer 1 1 TMA / test (6099) Gen-Probe - Aptima HPV Assay # * These tests exclusively detect "High Risk"-HPV (without specification of the HPV ) and cannot detect "Low Risk"-HPV. This means that a test result obtained by these tests can only be interpreted as " for High Risk-HPV". Results representing " for High Risk-HPV" will be stated as "" in your statement of individual. # This test system detects E6/E7 mrna of a distinct number of "High Risk" HPV s (see Table 3, Comment 3). 373 Human Papilloma Viruses Genome November December 2011 Table b.doc 12

19 Results for sample Differentiation of "High Risk" and "Low Risk" s of HPV Correct result: High Risk test in High Risk Low Risk indeterminate of High Risk Low Risk indeterminate of hybridization (70) Gen ID (AID) - HPV-screening Innogenetics - INNO-LiPA HPV Genotyp. Extra Gen ID (AID) - HPV-typing Greiner - PapilloCheck HPV Screening Roche - Linear Array HPV Genotyping Test in house Chipron - HPV Type nested PCR / hybridization (1270) in house Abbott - RealTime High Risk HPV hybridization / hybridization (3070) Qiagen - digene HC2 HPV DNA Test sequencing (72) in house nested PCR / sequencing (1272) in house test (99) PCR / test (1199) in house nested PCR / test (1299) in house Multiplex PCR / test (6199) manufacturer Human Papilloma Viruses Genome November December 2011 Table b.doc 13

20 Results for sample Typing of "High Risk" HPV and detection of "Low Risk" HPV (without typing) Correct result: HPV 18 test in HPV HPV 18 hybridization (70) Gen ID (AID) - HPV-screening Gen ID (AID) - HPV-typing Results for sample Typing of "High Risk" HPV and "Low Risk" HPV Correct result: HPV 18 test in HPV HPV 18 hybridization (70) Innogenetics - INNO-LiPA HPV Genotyp. Extra a, b Greiner - PapilloCheck HPV Screening Roche - Linear Array HPV Genotyping Test Chipron - HPV Type in house 5 5 c Gen ID (AID) - HPV-typing nested PCR / hybridization (1270) in house Abbott - RealTime High Risk HPV PCR / hybridization (9870) Sacace - HPV High Risk Typing a One participant detected additionally HPV 39. b One participant detected additionally HPV s 39, 68 and 73. c One participant detected additionally HPV Human Papilloma Viruses Genome November December 2011 Table b.doc 14

21 Results for sample Typing of "High Risk" HPV and "Low Risk" HPV (continued) HPV 18 sequencing (72) in house nested PCR / sequencing (1272) in house test (99) PCR / test (1199) in house Multiplex PCR / test (6199) manufacturer Results for sample Typing of "High Risk" HPV (without detection of "Low Risk" HPV) (Tests for exclusive detection and typing of "High Risk"-HPV )* Correct result: HPV 18 test in HPV 18 * HPV 18 * hybridization (70) Abbott - RealTime High Risk HPV sequencing (72) in house test (99) PCR / test (1199) AmpliSens - HPV-HCR-geno EPh TaqMan / test (1399) Roche - Cobas 4800 HPV Test manufacturer * These tests exclusively detect and "High Risk"-HPV and cannot detect "Low Risk"-HPV. This means that a test result obtained by these tests can only be interpreted as " for High Risk-HPV". Results representing " for High Risk-HPV" will be stated as "" in your statement of individual. 373 Human Papilloma Viruses Genome November December 2011 Table b.doc 15

22 Results for sample Detection of "High Risk" HPV (without detection of "Low Risk" HPV) (Tests which exclusively detect "High Risk"-HPV)* Correct result: High Risk test in High Risk Low Risk indeterminate of * High Risk Low Risk indeterminate of * hybridization (70) Roche - Amplicor HPV Test Abbott - RealTime High Risk HPV hybridization / hybridization (3070) Qiagen - digene HC2 HPV DNA Test sequencing (72) in house test (99) PCR / test (1199) manufacturer TaqMan / test (1399) manufacturer TMA / test (6099) Gen-Probe - Aptima HPV Assay # * These tests exclusively detect "High Risk"-HPV (without specification of the HPV ) and cannot detect "Low Risk"-HPV. This means that a test result obtained by these tests can only be interpreted as " for High Risk-HPV". Results representing " for High Risk-HPV" will be stated as "" in your statement of individual. # This test system detects E6/E7 mrna of a distinct number of "High Risk" HPV s (see Table 3, Comment 3). 373 Human Papilloma Viruses Genome November December 2011 Table b.doc 16

23 Results for sample Differentiation of "High Risk" and "Low Risk" s of HPV Correct result: High Risk test in High Risk Low Risk indeterminate of High Risk Low Risk indeterminate of hybridization (70) Gen ID (AID) - HPV-screening Innogenetics - INNO-LiPA HPV Genotyp. Extra Gen ID (AID) - HPV-typing Greiner - PapilloCheck HPV Screening Roche - Linear Array HPV Genotyping Test in house Chipron - HPV Type nested PCR / hybridization (1270) in house Abbott - RealTime High Risk HPV hybridization / hybridization (3070) Qiagen - digene HC2 HPV DNA Test sequencing (72) in house nested PCR / sequencing (1272) in house test (99) PCR / test (1199) in house nested PCR / test (1299) in house Multiplex PCR / test (6199) manufacturer Human Papilloma Viruses Genome November December 2011 Table b.doc 17

24 Results for sample Typing of "High Risk" HPV and detection of "Low Risk" HPV (without typing) Correct result: test in hybridization (70) Gen ID (AID) - HPV-screening Gen ID (AID) - HPV-typing Results for sample Typing of "High Risk" HPV and "Low Risk" HPV Correct result: test in hybridization (70) Innogenetics - INNO-LiPA HPV Genotyp. Extra Greiner - PapilloCheck HPV Screening Roche - Linear Array HPV Genotyping Test Chipron - HPV Type in house Gen ID (AID) - HPV-typing nested PCR / hybridization (1270) in house Abbott - RealTime High Risk HPV PCR / hybridization (9870) Sacace - HPV High Risk Typing Human Papilloma Viruses Genome November December 2011 Table b.doc 18

25 Results for sample Typing of "High Risk" HPV and "Low Risk" HPV (continued) sequencing (72) in house nested PCR / sequencing (1272) in house test (99) PCR / test (1199) in house Multiplex PCR / test (6199) manufacturer Results for sample Typing of "High Risk" HPV (without detection of "Low Risk" HPV) (Tests for exclusive detection and typing of "High Risk"-HPV )* Correct result: test in * * hybridization (70) Abbott - RealTime High Risk HPV sequencing (72) in house test (99) PCR / test (1199) AmpliSens - HPV-HCR-geno EPh TaqMan / test (1399) Roche - Cobas 4800 HPV Test manufacturer * These tests exclusively detect and "High Risk"-HPV and cannot detect "Low Risk"-HPV. This means that a test result obtained by these tests can only be interpreted as " for High Risk-HPV". Results representing " for High Risk-HPV" will be stated as "" in your statement of individual. 373 Human Papilloma Viruses Genome November December 2011 Table b.doc 19

26 Results for sample Detection of "High Risk" HPV (without detection of "Low Risk" HPV) (Tests which exclusively detect "High Risk"-HPV)* Correct result: High Risk test in High Risk Low Risk indeterminate of * High Risk Low Risk indeterminate of * hybridization (70) Roche - Amplicor HPV Test Abbott - RealTime High Risk HPV hybridization / hybridization (3070) Qiagen - digene HC2 HPV DNA Test sequencing (72) in house test (99) PCR / test (1199) manufacturer TaqMan / test (1399) manufacturer TMA / test (6099) Gen-Probe - Aptima HPV Assay # * These tests exclusively detect "High Risk"-HPV (without specification of the HPV ) and cannot detect "Low Risk"-HPV. This means that a test result obtained by these tests can only be interpreted as " for High Risk-HPV". Results representing " for High Risk-HPV" will be stated as "" in your statement of individual. # This test system detects E6/E7 mrna of a distinct number of "High Risk" HPV s (see Table 3, Comment 3). 373 Human Papilloma Viruses Genome November December 2011 Table b.doc 20