History of ILC Involvement in NAT Standardization

1. 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 • Industry Sponsored • ILC Members WHO, WHO Laboratories, International Regulatory Representatives • Presentation and discussion of design characteristics of Standard Reference Materials relevant to NAT Diagnostic Industry. • 2000: Development of International Standards: Stokes Court, UK • Industry Supported and Hosted: • ILC Members, WHO, WHO Laboratories, International Regulatory Representatives, Practicing Professionals (Physician and Laboratory) • How industry uses Standards. Specifications of appropriate standards for NAT diagnostics, impact of standardization on medical practice presented and discussed • Formation of a Working Party for WHO Meeting

2. History of ILC Involvement in NAT Standardization 2001 Planning Meeting for WHO Consultative Group Meeting: Lisbon Letter to WHO proposing Consultative meeting - EDMA endorsement of ILC as industry representative in this area 2002:WHO Consultation on International Standards for In Vitro Clinical Diagnostic of Viral Nucleic Acids WHO HQ, Geneva • WHO Sponsored • ILC Members, WHO, WHO Laboratories, International Regulatory Bodies, Practicing Professionals (Physician and Laboratory) • Design characteristics and requirements discussed • Outline of experimentation to test feasibility of synthetics standards • Formation of working group 2003: Phase I Experimentation and Report to SoGAT 2004: Additional Experimentation with newer tests • Proposal for further work by independent laboratories sponsored by industry.

3. Phase 1 Evaluation StudyObjectives • To compare quantification values for HCV RNA using in vitro-generated HCV RNA (quantified by analytical method) as calibrator. • Samples tested consisted of a panel of four dilutions of a patient-derived sample pool prepared by AcroMetrix (Benicia, CA) • Four different NAT methods were used to obtain these quantification values.

4. Phase 1 Evaluation StudyResults/Conclusions • The feasibility of using synthetic HCV RNA has been demonstrated • quantification values for members of the AcroMetrix panel were within 0.6 log10 • 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 HCV RNA transcript for quantification of international biological preparations are warranted. • determine further the feasibility of use of the HCV RNA transcript in different NAT technologies • develop quantification protocol that weighs data from different, validated methods or kits appropriately. • statistically-designed protocols that incorporate sources of variance

5. Phase 2 Evaluation StudyProposed Experiments • Reproducibility / Accuracy of the Dilutions of the Synthetic Standard Reference Material. • Gravimetric vs. volumetric dilutions with analysis in bDNA and TaqMan assays. • Reproducibility in Current Diagnostic / Quantitative Assays • Many replicates and runs of the diluted standard (diluted in the manner acceptable from the 1st experiment) in the following assays: • Abbott HCV RNA Assay (real-time PCR) • Bayer VERSANT HCV RNA 3.0 Assay (bDNA) • Roche HCV TaqMan Assay (real-time PCR)

6. Phase 2 Evaluation StudyProposed Implementation • ILC proposes that further work be carried out through an “independent” party, a reference or research laboratory that: • Can run all of the current and very near term commercial kits • Has demonstrated scientific acumen and objectivity.