EQASs for TTI Diagnostics as Tool for Elucidating and Solving Problems of Commercial and In-house Tests 21.03.2017 H. Zeichhardt1,3, V. Lindig1,2 and H.-P. Grunert1,2,3 1CharitéCentrum für diagnostische und präventive Labormedizin, Institut für Virologie, Campus Benjamin Franklin, Berlin 2Institut für Biotechnologische Diagnostik (GBD), Berlin 3INSTAND e.V., Düsseldorf Gesellschaft zur Förderung der Qualitätssicherung in medizinischen Laboratorien Collaborating Centers of International Consortium for Blood Safety (ICBS), New York Koch-Metchnikov-Forum of the Petersburg Dialogue Grunert, Bucharest, 2004

National and WHO International EQASs in Immunology/Virology 21.03.2017 National and WHO International EQASs in Immunology/Virology Influenzav. A/H5N1/B TBE Virus Rubellavirus Parvovirus B19 BSE (PrPsc) Herpes simpl. V. 2 Influenzav. A/H5N1/B Ag Hepatitis E Virus Herpes simpl. V. 1 Resp. Sync. V. Ag Hepatitis D Virus Varicella Zoster V. Mumpsvirus Hepatitis C Virus Cytomegalovirus Measlesvirus Hepatitis B V. Prg. II Adenoviruses HCV Genotyping Herpes simplex V. Hepatitis B V. Prg. I Enterov. (WHO/RKI) Hepatitis A Virus Enteroviruses Hepatitis B Virus Epstein-Barr Virus HIV-1 p24 Ag HPV HIV-1 (RNA) HIV-1/2 PCR/NAT Serology Samples total: > 702.000 (since 1988) Grunert, Bucharest, 2004

Partnership for National and International Quality Control Systems in Virus Diagnostics and Safety Testing of Blood 21.03.2017 CharitéCentrum für diagnostische und präventive Labormedizin Campus Benjamin Franklin, Institut für Virologie, Berlin Institute for Standardization and Documentation in Medical Laboratories (INSTAND), Düsseldorf WHO Collaborating Center for Quality Assurance and Standardization in Laboratory Medicine Institut für Biotechnologische Diagnostik der GBD, Berlin German Medical Association (Bundesärztekammer) German Association against Virus Diseases (DVV) Society of Virology (GfV) Paul Ehrlich Institute Robert Koch Institute National and International Reference Institutes Friedrich-Loeffler-Institute World Health Organization, Geneva and Lyon International Consortium for Blood Safety (ICBS), New York Diagnostic Council of DVV and GfV Grunert, Bucharest, 2004

INSTAND EQAS - Reference Laboratories 21.03.2017 Prof. Braun / Prof. Enders, Stuttgart Prof. Doerr / Prof. Rabenau / Dr. Berger, Frankfurt Prof. Fleckenstein / Dr. Huber / Dr. Korn, Erlangen Prof. Gerlich / Dr. Willems, Gießen Prof. Gürtler, Greifswald Dr. Heckler, Hannover Dr. Heim, Hannover Prof. Klenk / Prof. Radsak, Marburg Prof. Krüger / Dr. Meisel / Dr. Hofmann, Berlin Prof. Kurth / Prof. Pauli / Prof. Schreier / Dr. Mankertz / Dr. Schweiger / Dr. Tischer, RKI, Berlin Prof. Liebert / Prof. Pustowoit, Leipzig Prof. Löwer / Dr. Chudy / Dr. Nick / Dr. Nübling / Dr. Scheiblauer / Dr. Unger, PEI, Langen Prof. Mettenleiter / Prof. Groschup / Dr. Buschmann / PD Dr. Harder / Dr. Ziegler, FLI, Greifswald - Insel Riems Prof. Müller-Lantzsch / Dr. Gärtner, Homburg/Saar Prof. Neumann-Haefelin / Dr. Huzly, Freiburg Prof. Pfister / Dr. Wieland, Köln Prof. Roggendorf / PD Dr. Ross / Dr. Fiedler, Essen Dr. Vornwald, Berlin Prof. Wutzler / Prof. Faber / Dr. Sauerbrei, Jena Grunert, Bucharest, 2004

EQAS for Qualitative PCR/NAT HCV  Detection and solving of performance problems False negative results in inexperienced laboratories  Consequences: EQAS surplus samples for daily run controls and training

EQAS for HCV: Qualitative PCR/NAT (1) 21.03.2017 EQAS for HCV: Qualitative PCR/NAT (1) 99-100% 217 April 2002 183 November 2001 99% 206 April 2001 176 November 2000 201 April 2000 98-99% 171 November 1999 98-100% 194 April 1999 140 November 1998 92-99% New participants 156 April 1998 95-100% 103 November 1997 96-100% 102 July 1997 Correct results (for 4 samples) Participants Date Grunert, Bucharest, 2004

EQAS for HCV: Qualitative PCR/NAT (2) 21.03.2017 EQAS for HCV: Qualitative PCR/NAT (2) Results of new participants 91% (48/53) 53 new P 96% (99/103) 103 continuous P divided in 94% (147/156) 156 12004 87% (46/53) 94% (97/103) 92% (143/156) 12002 Apr. 1998 103 7 95% (98/103) 6 Nov. 1997 95% (97/101) 101 4 98% (100/102) 102 1 Jul. 1997 Correct results Participants (P) Sample No. Date Grunert, Bucharest, 2004

EQAS for Quantitative PCR/NAT HIV and HCV  Good lab performance with robust test: Benefit of EQAS samples run in dublicate in one scheme

Quantitative PCR/NAT Confidence Interval: Median +/- 0.5 log 21.03.2017 Quantitative PCR/NAT Confidence Interval: Median +/- 0.5 log Virus Date No. of samples Median (copies/ml) (IU/ml) Number of analyses Results in interval: median +/-0.5 log HIV-1 Nov 99-Mrz 07 Nov 02-Mrz 07 60 41 375-97,307 380-203,200 5419 282 93% (86%-99%) 82% (50%-100%) Grunert, Bucharest, 2004

EQAS for HIV-1: Lab Performance 21.03.2017 EQAS for HIV-1: Lab Performance Grunert, Bucharest, 2004

EQAS for HIV-1: Differentiation of Methods 21.03.2017 EQAS for HIV-1: Differentiation of Methods Grunert, Bucharest, 2004

Quantitative PCR/NAT Confidence Interval: Median +/- 0.5 log 21.03.2017 Quantitative PCR/NAT Confidence Interval: Median +/- 0.5 log Virus Date No. of samples Median (copies/ml) (IU/ml) Number of analyses Results in interval: median +/-0.5 log HIV-1 Nov 99-Mrz 07 Nov 02-Mrz 07 60 41 375-97,307 380-203,200 5419 282 93% (86%-99%) 82% (50%-100%) HCV*,& Apr 00-Mrz 07 57 54 685-1,351,081 673-400,000 1456 5050 75%*,& (27%-92%) 92%& (63%-100%) * without results for bDNA for samples <200,000 copies/ml (Nov. 1999 - Nov. 2000) & without results for bDNA for samples <3,000 copies/ml (Apr. 2004; Mrz. 2007) without results for bDNA for samples <1,220 IU/ml (Apr. 2004; Mrz 2007) Grunert, Bucharest, 2004

EQAS for Quantitative PCR/NAT HBV  Training with reliable tests: Benefit of identical EQAS samples for multiple testing

Improvement of Lab Performance by 2 Years Training 21.03.2017 Improvement of Lab Performance by 2 Years Training EQAS Nov 04, Nov 05 and Mar 07, HBV PCR/NAT (361) Copies/ml IU/ml Grunert, Bucharest, 2004

Quantitative PCR/NAT Confidence Interval: Median +/- 0.5 log 21.03.2017 Quantitative PCR/NAT Confidence Interval: Median +/- 0.5 log Virus Date No. of samples Median (copies/ml) (IU/ml) Number of analyses Results in interval: median +/-0.5 log HBV#,$,§ Nov 99-Mrz 07 Nov 01-Mrz 07 58 46 1,900-3,084,552 630-933,000 3281 1984 77%#,$ (52%-91%) 81%§ (55%-97%) # without results for bDNA and hybridization for samples <1,000,000 copies/ml (Nov. 1999 - Apr. 2003) $ without results for hybridization for samples <1,000,000 copies/ml (Nov. 2003 - Apr. 2004) § without results for bDNA for samples <500,000 IU/ml (Apr. 2002 - Apr. 2003) Grunert, Bucharest, 2004

EQAS for Quantitative PCR/NAT Cytomegalovirus  Performance of commercial and in-house tests

EQAS for CMV: Quantitative PCR/NAT 21.03.2017 EQAS for CMV: Quantitative PCR/NAT Date Participants (copies/ml) Correct results Median +/- 1.0 log (positive samples) Median +/- 0.5 log Nov 2001 30 70-80% 20-70% Apr 2002 43 88-93% 49-67% Nov 2002 40 78-88% 50-75% Apr 2003 52 90-96% 69-73% Nov 2003 50 92-94% 72-88% Apr 2004 59 90-93% 71-76% Nov 2004 58 90-95% 71-78% Apr 2005 66 94-97% 74-79% Aug 2005 24 92-96% 75-79% Nov 2005 67 91% 70-76% Mar 2006 37 81-100% 65-73% May 2006 65 89-94% Sep 2006 29 89-100% 72-82% Nov 2006 72 90-94% 64-72% Mar 2007 93-95% 81-88% Grunert, Bucharest, 2004

EQAS: Performance of Commercial and In-house Tests 21.03.2017 Grunert, Bucharest, 2004

Global Network for External Quality Control and Training 21.03.2017 External Quality Control (EQASs) EQASs (Germany, Europe, WHO) with >1075 laboratories with all routine tests up to 4-times per year Training Surplus samples from EQASs "Run" controls daily National reference materials "Run" controls in coop. with PEI Identical controls for:  Diagnostic labs  Blood banks  Industries  Licensing organizations (e.g. PEI)  Scientific labs Grunert, Bucharest, 2004

Contact Hans-Peter Grunert Heinz Zeichhardt 21.03.2017 Contact Hans-Peter Grunert Heinz Zeichhardt CharitéCentrum für diagnostische und präventive Labormedizin Institut für Virologie Campus Benjamin Franklin Hindenburgdamm 27 12203 Berlin Tel.: +49-30-8445 3624 or 3625 Fax: +49-30-8445 3626 Email: Hans-Peter.Grunert@charite.de Heinz.Zeichhardt@charite.de Grunert, Bucharest, 2004