Progress on Study to Validate Synthetic Materials for Use as Calibrators and References SoGAT XIX Bern, 14 June 2006 Roberta M. Madej Roche Molecular Diagnostics Representing the ILC

History of ILC Involvement in NAT Standardization 1998: Inter-Organization Discussion for the Establishment of Standard Reference Material for Nucleic Acid Diagnostics. Amsterdam, The Netherlands 2000: Development of International Standards: Stokes Court, UK 2001 Planning Meeting for WHO Consultative Group Meeting: Lisbon 2002: WHO Consultation on International Standards for In Vitro Clinical Diagnostic of Viral Nucleic Acids WHO HQ, Geneva 2003: Phase I Experimentation and Report to SoGAT 2004: Additional Experimentation with newer tests and dilution matrices 2005/ 2006: Redesign (and execution) of Phase II experiment ILC=Industry Liaison Committee – Active Molecular Test Manufacturers: Roche, Abbott, Bayer, BD, bioMerieux, Bio-Rad, Chiron, Gen-Probe

Phase 1 Evaluation Study Compared quantification values for HCV RNA using in vitro-generated HCV RNA (quantified by analytical method) as calibrator. –Four dilutions of a patient-derived sample pool prepared by AcroMetrix (Benicia, CA) –Four different NAT methods were used to obtain quantification values and recalculate based on synthetic calibrator values.

Phase 1 Evaluation Study Results/Conclusions The feasibility of using synthetic HCV RNA has been demonstrated quantification values for members of the AcroMetrix panel were within 0.6 log 10 greatest differences between quantification values from different NAT methods ranged between 0.39 and 0.57 for the 4 panel members Further studies examining the use of the synthetic material to contribute to calibration of continued lots of biologic standards. Determine feasibility of using the synthetic material in newer NAT assays Include more than one synthetic material Redesign experiment with equal comparability of all materials.

Phase 2 Evaluation Study “Parallel-line” experiment with –Synthetic RNA from Bayer –Asuragen armoured HCV RNA –PEI Reference (80,000 IU/ml) –HCV positive serum sample –HCV positive EDTA plasma sample –WHO Second International Standard for HCV (96/798, 100,000 IU/ml)** ( Materials “pre-diluted where possible.) Using the following Diagnostic / Quantitative Assays –Bayer VERSANT  HCV RNA 3.0 Assay (bDNA) /HCV RNA TMA QN Assay –Roche COBAS AmpliPrep  / COBAS TaqMan  HCV –Abbott RealTime TM HCV

Phase II Evaluation Study: Status Revised concept sent to outside reviewers (SoGAT members) Protocol edits in progress Materials: –Second synthetic material designed and trial samples sent to each participant –Sourcing of Basematrix –HCV High titre sample in progress Testing: –Asuragen test material: June / July –Actual Study: August-September –Data Analysis October

Phase II Evaluation Study: Participants Abbott: –G. Schneider Asuragen: –J. Hedges –E. Labourier –C. Walker –Peach Bayer: –S. Cruz –C. Bush-Donovan –J. Turczyn –J. Surtihadi Chiron: –B. Phelps Gen-Probe: –D. Kolk Roche: –R. Madej –J. Saldanha –Z. Wang ILC: –J. Reid