1 / 15

Performance of Chiron Quantitative and Qualitative HBV PCR Assay and Confirmation of HBV Yield Cases

June 13, 2007. Performance of Chiron Quantitative and Qualitative HBV PCR Assay and Confirmation of HBV Yield Cases. Yiu-Lian Fong, Ph.D, Associate Director. Agenda.

glynn
Download Presentation

Performance of Chiron Quantitative and Qualitative HBV PCR Assay and Confirmation of HBV Yield Cases

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. June 13, 2007 Performance of Chiron Quantitative and Qualitative HBV PCR Assay and Confirmation of HBV Yield Cases Yiu-Lian Fong, Ph.D, Associate Director

  2. Agenda A qualitative and quantitative HBV DNA assay has been developed and validated in house for sensitive detection and accurate quantification of HBV DNA for confirmatory purposes. • Assay Performance • Confirming HBV Yield Cases • Issues raised for standardized calibration of various genotypes

  3. Assay Performance • Analytical Sensitivity and LOD for WHO HBV Standard • Analytical Sensitivities for DDL Genotypes A-G • Linearity, LLOQ, reproducibility and precision • Clinical Sensitivity • Analytical and Clinical Specificities

  4. Analytical Sensitivity and LOD for WHO HBV Standard % Positive HBV (IU/mL)

  5. Analytical Sensitivity and LOD for WHO HBV Standard *Procleix Ultrio PI, IN0166EN Rev. A

  6. Analytical Sensitivity for DDL HBV Genotypes

  7. Reproducibility, Linearity, Accuracy, and LLOQ R2 = 0.995 - 0.999 CV = 0.4% - 1.3% LLOQ = 3.3 IU/mL

  8. 15 days Clinical Sensitivity: Early Detection of Seroconversion Panel #6289 *Acrometrix PI. **Combined testing results from multiple sites.

  9. 23-28 days Clinical Sensitivity: Early Detection of Seroconversion Panel #11006 *Acrometrix PI. **Combined testing results from multiple sites.

  10. Clinical Sensitivity: Early Detection of Seroconversion Panel #11008 18-33 days *Acrometrix PI. **Combined testing results from multiple sites

  11. Analytical and Clinical Specificities In-house Assay Validation: • 100 random donor samples tested in triplicates: all negative • Freeze/thaw up to five cycles with 5, 20, and 5000 IU/mL WHO HBV standard • Potential Interfering Substances Tested • Anticoagulants: ACD, CPD, K2EDTA, K3EDTA, Na Citrate, Na Heparin • Protein at > 7 g/dL • Lipid/Triglyceride at > 3000 mg/dL • Hemoglobin at > 500 mg/dL • Bilirubin at >20 mg/dL • Bacteria (>105 CFU/mol) commonly seen in blood related products • Blood-born pathogens:HCV, HTLV, HAV, CMV, EBS, HSV, and HIV-1

  12. Confirming HBV Yield Cases: Chiang Mai University Study (HBsAg Negative)* *Nantachit, N. et al, in press, 2007. **Later HBsAg positive. nd, not determined.

  13. Quantitation of DDL Genotype A-G * Supplied as 30,000 Cp/mL.

  14. Summary • A sensitive qualitative and quantitative HBV DNA assay has been developed and validated in house • LOD at 3.3 IU/mL for WHO HBV standard • Detects various genotypes • Detects window phase low viremia samples • Confirms yield cases • Conclusion: This highly sensitive and reliable assay is suitable for confirmatory purposes • Remaining Issues: • How to standardize? • How to calibrate various genotypes? • Which test method to use?

  15. Acknowledgement • Dennis Madriaga • Baohe Shen • Barney Krebs

More Related