1. HIV, HCV, and HBV NAT Controls Formulation, Stability and Performance Mark Manak BBI Diagnostics, Inc. A Division of SeraCare Life Sciences, Inc. SoGAT XX Meeting Warsaw, Poland June 12-13, 2007

2. Quantitative viral load testing is used for monitoring disease progression, or response to therapy Need for reliable quantitation data to ensure proper patient management Report of the NIH Panel to Define Principles of Therapy of HIV Infections and Guidelines of the Use of Antiretroviral Agents in HIV Infected Adults and Adolescents. MMWR 1998:46 (no.RR-5) A log 0.5 increase in HIV titer in two sequential samples is considered a significant increase. Need for Precision of Quantitation

3. Viral Load False Positive Rate vs. Interassay SD % False Positive Rate Biological Variation SD 0.24 log (10) 3% of patients will falsely be classified as having significantly increased viral load Standard Deviation Small increases in interassay variation significantly affect both false poistive and false negative rates Assays are complex, many factors may negatively affect interassay variation

4. HIV, HBV, and HCV ACCURUN Controls Manufactured from whole virus Non-infectious cultured HIV (8E5), or HBV or HCV positive plasma Diluted into Defibrinated Plasma (Basematrix) Resemble patient specimen Different Series target different viral load levels Manufactured under cGMP and ISO 13485 FDA and CE Mark

5. Independent Run Control Independent of the test kit (manufacurer, reagents, calibration) Designed to perform with a wide number of test kits and/or procedures Control limits are usually set by the individual laboratory Intended Use – Estimate laboratory testing precision – Detect errors in laboratory testing procedures. – Controls for the entire workflow

6. +3SD +2SD Mean -2SD -3SD Levy Jennings Plot of A315 Values Log IU/ml Date

7. Consistency of A315 HIV Controls IU/ml

8. Real Time Stability of 8E5 Stock IU/ml

9. Sources of Variability HIV NAT IU/ml

10. Consistency of HCV NAT: Effect of Kit Lot and A325 Lot ACCURUN Kit Lot IU/ml

11. Real Time Stability HCV A325 S300

12. Consistency of HBV NAT: Effect of Kit Lot and A345 Lot

13. Quantitation Controls/Panels Available ACCURUN Controls – HIVA315 Series 100, 200, 300, 400, 500 – HCV A305 Series 100, 200, 300, 400 A405 Series 100 calibrated to IU – HBV A325 Series 300, 600, 700 Linearity Panels – Series of 10 Dilutions ranging from 3 x 10 6 – 10 IU/ml – HIV Linearity Panel PRD801 – HCV Linearity Panel PHW804 – HBV Linearity Panel PDH801

14. ACCURUN Controls and Panels Stable, independent quality control material Made from Whole virus in Defibrinated Plasma Resemble patient specimen Long term stability when stored frozen at -70 o C Controls for all test steps: extraction, amplification and detection Reliable Performance Manufactured under audited quality system Targeted to decision levels Large lot sizes to enable usage over extended periods Excellent Reproducibility between Control Lots, Test Runs, and Test Sites Access to technical support

15. Acknowledgements SeraCare –Debbie Petit –Pat Garrett –Bharathi Anekella –Joan Thurston Quest Diagnostics Quintiles Dallas VA Medical Center NIH Clinical Lab Medical Center of New Orleans SeraCare Testing Lab