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ICBS Master Panels for Kit Evaluation: HCV, HBV, and HIV (WWW.ICBS-WEB.ORG)

ICBS Master Panels for Kit Evaluation: HCV, HBV, and HIV (WWW.ICBS-WEB.ORG). Howard A. Fields, Ph.D. Susan Diaz, MPH Division of Viral Hepatitis Centers for Disease Control and Prevention. Objective of Master Panels.

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ICBS Master Panels for Kit Evaluation: HCV, HBV, and HIV (WWW.ICBS-WEB.ORG)

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  1. ICBS Master Panels for Kit Evaluation: HCV, HBV, and HIV(WWW.ICBS-WEB.ORG) Howard A. Fields, Ph.D. Susan Diaz, MPH Division of Viral Hepatitis Centers for Disease Control and Prevention

  2. Objective of Master Panels • Identify, validate, and make available high-quality affordable tests for HBV, HCV, and HIV because cost is one of the major impediments to universal screening. • Use as a model for BTEP project to produce Master Panels with Dr. Alexander Kanev, Vector currently in year 3.

  3. HCV Master Panelspositive and negative

  4. HCV Master Panel:Diversity • Country of origin • Genotype • Subtype • RIBA 3.0 banding patterns

  5. * N=14 for HCV some collaborators different for HBV and HIV Location of International HCV Collaborators

  6. Methods to Characterize Panel Members • Screening Tests • Anti-HCV by Ortho 3.0; positives repeated in duplicate • Confirmatory Tests • RIBA 3.0 for HCV by Chiron • Quantitative PCR • Roche Amplicor for HCV RNA • Genotyping / Subtyping by direct DNA Sequencing • CDC: ABI Prism 3100 Capillary Sequencer (NS5b default to 5’-UTR); phylogenetic analysis • Visible Genetics, Inc. Clip Sequencing Reaction for HCV (5’UTR/NS5b) percent homology

  7. Collections • HCV: 1007 • HBV: 450 • HIV: 200 • COINFECTION: 62 • NEGATIVES: 450 • Total Units Received: 2169

  8. HCV Positive UnitsDescription of Selectable Units • Serologically confirmed as positive (exclude indeterminate specimens) • Concordant genotype by direct DNA sequencing from two laboratories • Minimum volume 100 ml • Additional selection criteria based upon diversity • By geographic regions • By genotype/subtype • By RIBA banding patterns

  9. Diversity by Genotype among Selected Units • Among 1007 HCV IR units received and characterized, 539 unitsmet the definition of selectable (54%) • Genotype 1 (n=296) • Genotype 2 (n=26) • Genotype 3 (n=42) • Genotype 4 (n=129) • Genotype 5 (n=3) • Genotype 6 (n=43)

  10. 66 23 40 36 3 32 Diversity by subtype RIBA 3.0 banding patterns Diversity by Country and Genotype among Official HCV Panel Members

  11. Negative Panelgeographic diversity • Manufacturing completedJune 18, 2002 • First panel to be completed • 200 Members from both thedeveloped and developing world • 181 domestic members utilized from ARC • 19 members pooled by region: Egypt, Indonesia, and Ivory Coast

  12. Origin of Kits Sent to PEI • China 13 • Russia 9 • India 7 • South Korea 5 • EU 4 • Costa Rica 1 • Indonesia 1 • Uzbekistan 1 • Brazil 1 • Argentina 1 • USA (not FDA approved) 1 Total = 44

  13. Russian Kits Sent to PEI for Evaluation • Biotechnologitzeskaja Kompanija Bioservis (No. 23, Hepascreen) • IMBIO (No. 24, Anti-HCV ELISA) • Medizinische Biologische Union (No.27, HCV-DCM) • Union–Diagnostisches-System (No.25, Anti-HCV ELISA) • Vector-Best (No. 29, RecombiBest anti-HCV-Strip, OPD) • Received, but Not Evaluated as of August 23, 2004 • Institut Pastera Sankt-Peterburg (Anti-HCV ELISA-G) • NPP Akwapast (Immunoenzyme Test-System for Detection of Anti-HCV) • Radiopreparat (Anti-HCV) • Vector-Best (RocombiBest anti-HCV-Strip, TMB)

  14. Specimens • ICBS HCV Master Panel (n=200) • ICBS negative Master Panel (n=181 and 200) • Seroconversion Panel (commercial) • core only, genotype 2b • NS3 only • Core & NS3, genotype 1a • Mean day delay

  15. ^ ^ ^ ^ ^ * * * * * ^ Russian kits * Significant reduction in specificity (n = 181 vs. 200)

  16. * * * * * ^ ^ ^ ^ ^ ^ Russian kits * Significant reduction in specificity (n = 181 vs. 200)

  17. * * PI of Russian Kits Evaluated PI • Union–Diagnostisches-System235.28 232.83 • Biotech Kompanija Bioservis235.00 231.00 • IMBIO 232.83 232.33 • Vector-Best 231.78 229.83 • Medizinische Biolog Union 231.28 229.30 N=181 N=200 * Statistical significant reduction in specificity (n = 181 vs. 200)

  18. Conclusions (1) • ICBS HCV Negative Master Panel manufactured June, 2002. • N = 181 US specimens • N =19 specimens from the developing world yielded weaklyscreening test positive when tested with some assays licensed in the EU. Some specimens also tested positive/indeterminate with some of the EU license supplemental tests. • N = 200 specimens confirmed as true negative at CDC(Ortho 3.0, Chiron RIBA 3.0, and Roche HCV Amplicor)

  19. Conclusions (2) • ICBS HCV Positive Master Panel manufactured September, 2002 • Diversity based on geographic origin, genotype, subtype (concordance by DNA sequencing), and RIBA 3.0 banding patterns • All confirmed as true positive by screening (Ortho 3.0), supplemental testing (RIBA 3.0), and positive PCR (Roche HCV Amplicor)

  20. Conclusion (3) • Russian kits evaluated by PEI 2003 – 2004 • PI less than kits licensed in the US and EU and several other kits manufactured in resource-challenged regions of the world • Highest PI manufactured by Union–Diagnostisches-Systemprobably equal to the kit manufactured by Biotech Kompanija Bioservisbased on 181US specimens • When 19 specimens from the developing world was added (n=200), the specificity of all 5 Russian kits decreased; however, only kits Union–Diagnostisches-System and Biotechnologitzeskaja Kompanija Bioservis decreased to a statistical significant level. • Despite this decrease in specificity, the kit manufactured by Union–Diagnostisches-System continued to have the highest PI value among the Russian kits evaluated.

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