1 / 16

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

hal
Download Presentation

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

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  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

More Related