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Linearity Panels HIV RNA, HCV RNA, HBV DNA, and CMV DNA

Linearity Panels HIV RNA, HCV RNA, HBV DNA, and CMV DNA. Mark Manak, Ph.D. BBI Diagnostics, Inc. A Division of SeraCare Life Sciences, Inc. SoGAT XVIII Bethesda, MD May 24-25, 2005. Linearity Panel Design. Mimic clinical specimen Whole virus in defibrinated plasma diluent

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Linearity Panels HIV RNA, HCV RNA, HBV DNA, and CMV DNA

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  1. Linearity PanelsHIV RNA, HCV RNA, HBV DNA, and CMV DNA Mark Manak, Ph.D. BBI Diagnostics, Inc. A Division of SeraCare Life Sciences, Inc. SoGAT XVIII Bethesda, MD May 24-25, 2005

  2. Linearity Panel Design • Mimic clinical specimen • Whole virus in defibrinated plasma diluent • Challenges the entire analytical process • One negative, and Nine positive members • Series of sequential one log or half log dilutions • Results expressed as specified by kit manufacturers • Serial dilutions of the WHO International Standards analyzed simultaneously to assign IU values, where appropriate

  3. Intended Use Assay Effectiveness • Evaluate sensitivity and dynamic range of quantitative viral nucleic acid assays • Identifying consistency over a broad linear range • Establish calibrations vs. IU Standards • Verification of performance at kit lot changes Training • Operator training program • Operator validation Part of Laboratory Quality Assurance program

  4. HCV RNA Linearity Panel (PHW804) Diluent: • Normal human plasma • Negative for antibody to HIV-1/2, HBsAg, HCV, HTLV-I/II • Negative for HBsAg, HIV-1 RNA, HBV RNA and HCV DNA Virus Spike: • High titer HCV RNA positive human plasma • HCV Genotype 1 by Innogenetics InnoLIPA Assay Results on Data Sheet: • Roche COBAS Amplicor HCV Monitor v2.0 • Bayer Versant HCV 3.0 bDNA Assay • Abbott LCx HCV Quantitative Assay • QuantaSure Plus HCV Roche Taqman ASR

  5. HCV RNA Unit Conversions to IU • Roche COBAS: 2.2 copies/IU • Bayer Versant: 5.2 copies/IU • Abbott LCx: 4.3 copies/IU • Taqman: 0.6 copies/IU

  6. HIV RNA Linearity Panel (PRD801) Diluent: • Normal human plasma • Negative for anti-HIV-1/2, anti-HBsAg, anti-HCV, anti-HTLV-I/II • Negative for HBsAg, HIV-1 RNA, HBV RNA and HCV DNA Virus Spike: • Culture supernatant: HIV-1 Type B 8E5 virus • Intact, but defective virus genome Results on Data Sheet: • Roche COBAS Amplicor HIV Monitor v1.5 • Roche COBAS Amplicor HIV Monitor v1.5 Ultra • Bayer Versant HIV-1 RNA bDNA 3.0 Assay • Chiron/Genprobe Procleix HIV-1 RNA Discriminatory Qualitative • BioMerieux Nuclisens HIV-1 QT

  7. HBV DNA Linearity/Sensitivity Panel (PHD801) Diluent: • Normal human plasma • Negative for antibody to HIV-1/2, HBsAg, HCV, HTLV-I/II • Negative for HBsAg, HIV-1 RNA, HBV RNA and HCV DNA Virus Spike: • High titer HBV DNA positive human plasma • HBV DNA:Type A virus by Roche Taqman and Innogentics InnoLIPA • Serotype ad Results on Data Sheet: • Roche COBAS Amplicor HBV • Digene HBV DNA Quantitative Assay • Bayer Versant HBV DNA Quantitative bDNA Assay • Taqman HBV DNA Quantitative Assay

  8. HBV DNA Unit Conversions to IU • Roche COBAS: 1.2 copies/IU • Bayer Versant: 0.9 copies/IU • Taqman: 3.9 copies/IU • Digene: 4.6 copies/IU • Digene: 1,200,000 IU/pg

  9. CMV DNA Linearity/Sensitivity Panel (PTC801) Diluent: • Normal human plasma • Negative for antibody to HIV-1/2, HBsAg, HCV, HTLV-I/II • Negative for HBsAg, HIV-1 RNA, HBV RNA and HCV DNA • Negative for anti-CMV and CMV DNA Virus Spike: • Culture supernatant from CMV cultures Results on Data Sheet: • Roche COBAS Amplicor CMV Monitor Test

  10. Conclusions BBI Linearity panels are made from natural virus in plasma diluent Nine positive members spanning a broad range of virus concentrations and a negative member Calibrated against WHO standards and expressed in IU/ml Data from available commercial assays Useful for monitoring linearity and dynamic range of assays

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