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Update on the Evaluation of the Behavior of Synthetic Materials and Biologics. SoGAT XX Warsaw, 12 June 2007 Roberta M. Madej Roche Molecular Diagnostics Representing the ILC. Purpose.
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Update on the Evaluation of the Behavior of Synthetic Materials and Biologics SoGAT XX Warsaw, 12 June 2007 Roberta M. Madej Roche Molecular Diagnostics Representing the ILC
Purpose • To evaluate the behavior of synthetic materials and biologics in three commercially available quantitative HCV NAT assays.
Design “Parallel-line” experiment with • Synthetic RNA from Siemens • Asuragen armored 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 RealTimeTM HCV Six dilution points of five different materials in three different assays.
Considerations • Materials • Asuragen and Siemens Materials • Donated by Asuragen and Siemens • Calibrated by independent means to copies/ml Phosphate std and OD readings • PEI material • Donated by M. Nübling • Calibrated to the WHO material std in IU/ml (80,000 IU/ml) • Plasma and Serum • Made and donated by Abbott • Initial testing by Abbott • Assays and logistics • Siemens assay is calibrated with the Siemens standard material already and reports in copies/ml • Abbott and Roche: calibrated against WHO standard: report in IU/ml • 5 copies/IU used throughout the study • PEI and Siemens material diluted at each site into the same base matrix. (PEI was also reconstituted at each site) • Asuragen, Plasma and Serum were diluted and tested BEFORE distribution to each site.
Comparative Characteristics • Within Run cv’s 4-60% • Across Run cv’s: 8-46% • Across Platform cv’s: 19-117%
Variability Gage Charts Asuragen Siemens Plasma PEI
Summary • The synthetic materials behaved in a similar manner to the biologic materials in the three assays used. • Materials were recovered by the three assays used. • Parallel along the linear range • Similar variability • Relative quantification of each material was not necessarily consistent among the methods.
Biologic Standard Synthetic Is There a Role? Can we utilize the attributes of both types of materials to maximize the information? Establish Test Test Test Test
Issues to be addressed • Stability • Manufacturability • Commutability • Science is moving forward: • Whole length genome? • Cultured virus? • Subject to the same questions
Abbott: G. Schneider G. Gabriel Asuragen: J. Hedges E. Labourier C. Walker Peach PEI Micha Nübling Roche: R. Madej J. Saldanha Z. Wang G. Schilling C. Calado Siemens: S. Cruz A. Ocampo ILC (Industry Liaison Council) J. Reid (Chair) Abbott G. Schneider BioMerieux P. van de Weil R. Bally Bio-Rad M. Gonzales J. Jones A. Khellaf Chiron-Novartis N. Lelie Gen-Probe D. Kolk Roche P. Colucci R. Madej Siemens J. Turczyn Evaluation Study: Participants