AREVIR-GenaFor-Meeting Bonn, 05.05.11 ________________ Comparison - PowerPoint PPT Presentation

AREVIR-GenaFor-Meeting Bonn, 05.05.11 ________________ Comparison of HIV-1 Viral Load samples between Real Time (Abbott) and COBAS TaqMan (Roche) ________________ Gudrun Naeth PZB AACHEN Blondelstr. 9 52062 Aachen

Introduction Introduction � Low Level Viremia � Comparision of Roche COBAS TaqMan vs. Abbott m2000 � Results PZB PZB PZB AACHEN AACHEN AACHEN

Transient low low level level viremia viremia during during ART ART Transient date ART HIV-1 RNA CD4 absolute CD4 relative 01.2001 AZT 3TC EFV 11.2001 AZT 3TC LPV/r 03.2002 <50 643 33 01.2003 TDF 3TC LPV/r 04.2004 <50 03.2005 47 01.2006 01.2007 <40 (det.) 641 37 01.2008 <40 (non det.) 617 31 02.2009 <40 (det.) 715 33 05.2009 59 617 31 07.2009 <40 641 37 11.2009 72 675 38 01.2010 40 659 37 PZB PZB PZB 05.2010 71 615 37 AACHEN AACHEN AACHEN 69

Current Treatment Treatment Recomendations Recomendations Current goal of therapy virologic failure >50 copies/ml Germany 1 <20-50 copies/ml „evaluate virologic failure“ Europe 2 <50 copies/ml 2x >50 copies/ml 2x >50 copies/ml (PCR) International 3 <50 copies/ml (PCR) 2x >75 copies/ml (bDNA) <75 copies/ml (bDNA) International 4 <48 copies/ml 2x >200 copies/ml 1 Deutsche AIDS-Gesellschaft e.V. ( DAIG ). Deutsch-Österreichische Leitlinien zur Antiretroviralen Therapie der HIV-1-Infektion (04.02.2010). PZB PZB PZB 2 European AIDS Clinical Society ( EACS ). Guidelines. Clinical Management and Treatment of HIV-infected Adults in Europe. Version 5-3. 2011. 3 International AIDS Society ( IAS ). Recommendations. Antiretroviral Treatment of Adult HIV Infection. 2010. AACHEN AACHEN AACHEN 4 Department of Health and Human Services ( DHHS ). Guidelines for the Use of Antiretroviral Agents in HIV-1 infected Adults and Adolescents (2011).

HIV- -viral viral load load measurements measurements and and resistance resistance analysis analysis HIV (PZB Aachen) (PZB Aachen) 40-100 copies/ml: 4,2% 101-1000 copies/ml: 5,9% >1000 copies/ml: 17,8% number of measurments 7.719 66,2 % 63,7% 61,2 % 61,1 % <40 <40 <50 <40 71,6% cop/ml cop/ml cop/ml cop/ml <40 Kop/ml n=910 n=1.243 n=1.353 28.06.2010 PZB PZB PZB Genotypic resistance analysis: reverse transcriptase and protease AACHEN AACHEN AACHEN Braun P., 13 MAT, 2010

Therapy success success = = viral viral load load below below detection detection limit limit?? ?? Therapy 1.000.000 1.000 detection limit Abbott RealTime: 40 copies/ml 100.000 800 Roche TaqMan: 20 copies/ml HIV RNA [Kopien/ml] 10.000 Siemens Versant kPCR: 37 copies/ml CD4 [Zellen/µl] 600 1.000 400 100 200 10 1 0 Nov 07 Jan 08 Feb 08 Mrz 08 Apr 08 Jun 08 Jan 09 Jan 10 Jan 11 antiretroviral therapy PZB PZB PZB AACHEN AACHEN AACHEN

Abbott m2000 / Roche TaqMan TaqMan v 1.0 v 1.0 Abbott m2000 / Roche Abbott Real Time HIV-1 assay Cobas Ampli-Prep Cobas TaqMan HIV-1 assay Automated m2000sp Automated COBAS AmpliPrep m2000rt COBAS TaqMan Pol IN Gag 0,6 mL 1,0 mL 40-10.000.000 copies/mL 20-10.000.000 copies/mL PZB PZB PZB AACHEN AACHEN AACHEN

Reproducibility and Precision Two real-time PCR assays with international standards and serial dilutions of a patient sample PEI-Standard (Paul-Ehrlich-Institut) • 40,000 IU/ml = 24,000 copies/ml • Three dilutions: 1x10 2 , 5x10 2 , and 1x10 3 copies/ml • 20-fold measurement WHO-Standard (World-Health-Organisation) • 100,000 IU/ml = 60,000 copies/ml • Five dilutions: 1x10 2 , 2.5x10 2 , 5x10 2 , 1x10 3 , and 2.5x10 3 copies/ml • 20-fold measurement for lowest viral load and 10-fold for other dilutions Patient Sample (PZB Aachen) • Seven dilutions: 5x10 1 , 1x10 2 , 1x10 3 , 1x10 4 , 5x10 4 , 1x10 5 and 5x10 5 copies/ml • 10-fold measurement PZB PZB PZB AACHEN Braun P., Ehret R., Wiesmann F, et al. Comparison of four commercial quantitative HIV-1 assays. Clin Chem Lab Med; 2007; 45: 93-99 AACHEN AACHEN

Reproducibility and Precision PEI-Standard 20-fold measurement m1000/m2000rt CAP/CTM v 1.0 A B R 2 =0,91 R 2 =0,72 16% CV 41% CV 49% CV 16% CV 32% CV 56% CV 5x10 2 1x10 3 5x10 2 1x10 3 1x10 2 1x10 2 PZB PZB PZB AACHEN Braun P., Ehret R., Wiesmann F, et al. Comparison of four commercial quantitative HIV-1 assays. Clin Chem Lab Med; 2007; 45: 93-99 AACHEN AACHEN

Reproducibility and Precision WHO-Standard 20 (10)-fold measurement m1000/2000rt CAP/CTM v 1.0 C D R 2 =0,96 R 2 =0,88 Measurement [copies/ml] Measurement [copies/ml] 25% CV 14% CV 47% CV 21% CV 15% CV 20% CV 12% CV 22% CV 23% CV 48% CV 1x10 2 2,5x10 2 5x10 2 1x10 3 1x10 2 2,5x10 2 5x10 2 1x10 3 2,5x10 3 2,5x10 3 Nominal level [copies/ml] Nominal level [copies/ml] PZB PZB PZB AACHEN Braun P., Ehret R., Wiesmann F, et al. Comparison of four commercial quantitative HIV-1 assays. Clin Chem Lab Med; 2007; 45: 93-99 AACHEN AACHEN

Reproducibility and Precision Clinical Sample m2000rt m2000rt TaqMan v 1.0 v 1.0 TaqMan 1000000 Measurement [copies/ml] 100000 19% CV 34% CV 10% CV 28% CV 10000 9% CV 23% CV 36% CV 18% CV 1000 11% CV 39% CV 100 37% CV 18% CV 10 5,0 5,7 5,0 5,7 PZB 1,7 2,0 3,0 4,0 4,7 PZB 1,7 2,0 3,0 4,0 4,7 PZB Expected concentrations in log copies/ml AACHEN AACHEN AACHEN

Correlation on Clinical Samples m2000rt versus TaqMan v 1.0 R 2 : 0,937 m2000rt [log copies/mL] TaqMan minus m2000rt Differences (mean) in log 10 copies/mL = 0,22 PZB PZB PZB TaqMan [log copies/mL] AACHEN Braun P., Ehret R., Wiesmann F, et al. Comparison of four commercial quantitative HIV-1 assays. Clin Chem Lab Med; 2007; 45: 93-99 AACHEN AACHEN

Evaluation: Low level level viremia viremia Evaluation: Low Comparison between Roche TaqMan (version 2) and Abbott m2000 Slides not available Results in press, 2011 PZB PZB PZB AACHEN AACHEN AACHEN

Retrospective analysis of plasma samples (detectable and undetectable viral load) with RealTime and TaqMan v. 2.0 5 Groups (10 patients, 5 measuring points) Group 1: VL <40 copies/ml (not detected) Group 2: VL <40 copies/ml (detected not + detected) Group 3: VL <40 copiesn/ml (not detected) Group 4: VL >40 – 100 copies/ml Group 5: VL >40 – 200 copies/ml PZB PZB PZB AACHEN AACHEN AACHEN

Retrospective Analysis Example Date Previous value m2000 CTM 24.08.2010 41 63 116 17.08.2009 87 87 206 6 19.02.2009 79 73 192 20.10.2008 193 91 219 18.07.2008 120 97 168 PZB PZB PZB AACHEN AACHEN AACHEN

Thank you for your attention! PZB PZB PZB AACHEN AACHEN AACHEN