![Page 1: 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](https://reader035.vdocuments.us/reader035/viewer/2022062320/56649d0a5503460f949dc490/html5/thumbnails/1.jpg)
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 XVIIIBethesda, MD
May 24-25, 2005
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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
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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
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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
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HCV Linearity Panel (PHW804)
10
100
1,000
10,000
100,000
1,000,000
10,000,000
1101001,00010,000100,0001,000,000
Dilution
IU/m
l
Roche COBAS
Bayer Versant
Abbott LCx
Roche Taqman
Power (RocheTaqman)Power (AbbottLCx)Power (RocheCOBAS)Power (BayerVersant)
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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
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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
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HIV RNA Linearity Panel (PRD801)
10
100
1,000
10,000
100,000
1,000,000
10,000,000
1101001,00010,000100,000
Dilution
Cop
ies/
ml
Roche COBAS
Roche COBAS Ultra
Bayer Versant
Nuclisens
Power (Roche COBASUltra)Power (Nuclisens)
Power (Nuclisens)
Power (Nuclisens)
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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
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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/IUDigene: 1,200,000 IU/pg
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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
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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