Скачать презентацию June 13 2007 Performance of Chiron Quantitative and Скачать презентацию June 13 2007 Performance of Chiron Quantitative and

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June 13, 2007 Performance of Chiron Quantitative and Qualitative HBV PCR Assay and Confirmation 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 A qualitative and quantitative HBV DNA assay has been developed and validated in 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 2 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only)

Assay Performance § Analytical Sensitivity and LOD for WHO HBV Standard § Analytical Sensitivities 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 3 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only)

% Positive Analytical Sensitivity and LOD for WHO HBV Standard HBV (IU/m. L) ED % Positive Analytical Sensitivity and LOD for WHO HBV Standard HBV (IU/m. L) ED 95 HBV # Positive % IU/m. L / # Tested Positive 20 20/20 100 5 20/20 100 2. 50 18/20 90 1. 25 13/20 65 0. 63 8/20 40 0. 31 3/20 15 0 0/16 0 3. 26 (2. 21 – 6. 53) IU/m. L 4 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only)

Analytical Sensitivity and LOD for WHO HBV Standard Expected Dose (range), IU/m. L Detection Analytical Sensitivity and LOD for WHO HBV Standard Expected Dose (range), IU/m. L Detection Rate Chiron QPCR Ultrio* Ultrio d. HBV* ED 95 3. 3 (2. 2– 6. 5) 8. 0 (7. 1– 9. 3) 6. 8 (6. 0– 7. 7) ED 50 0. 8 (0. 6– 1. 0) 3. 3 (3. 0– 3. 8) 3. 0 (2. 7– 3. 4) *Procleix Ultrio PI, IN 0166 EN Rev. A 5 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only)

Analytical Sensitivity for DDL HBV Genotypes HBV Genotype Genotype Cp/m. L A B C Analytical Sensitivity for DDL HBV Genotypes HBV Genotype Genotype Cp/m. L A B C D E F G 1000 100 100 300 100 100 100 100 50 100 75 82 83 100 30 100 75 92 64 50 83 100 10 92 33 33 8 17 42 92 3 27 33 33 20 33 17 90 0 0 0 0 6 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only)

Reproducibility, Linearity, Accuracy, and LLOQ R 2 = 0. 995 - 0. 999 CV Reproducibility, Linearity, Accuracy, and LLOQ R 2 = 0. 995 - 0. 999 CV = 0. 4% - 1. 3% LLOQ = 3. 3 IU/m. L 7 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only)

Clinical Sensitivity: Early Detection of Seroconversion Panel #6289 Day s *ABBOTT PRISM HBs. Ag Clinical Sensitivity: Early Detection of Seroconversion Panel #6289 Day s *ABBOTT PRISM HBs. Ag **Ultrio #Positive / #Tested 0 0. 95 2 Chiron QPCR Results #Positive / #Tested IU/m. L 0/16 0/3 NR 0. 20 0/17 0/3 NR 9 0. 56 0/17 2/3 11 0. 50 2/18 0/3 16 0. 21 12/17 3/3 18 0. 22 16/17 3/3 *Acrometrix PI. **Combined testing results from multiple sites. 31 2. 15 18/18 3/3 8 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only) 36 4. 94 18/18 3/3 < 3. 3 NR 6. 2 14. 1 962. 9 6212. 15 days

Clinical Sensitivity: Early Detection of Seroconversion Panel #11006 Day s *ABBOTT PRISM HBs. Ag Clinical Sensitivity: Early Detection of Seroconversion Panel #11006 Day s *ABBOTT PRISM HBs. Ag **Ultrio #Positive / #Tested 0 0. 26 0/17 0/5 3 7 10 0. 16 0. 27 0. 24 0/18 0/16 0/5 0/5 14 0. 24 2/18 2/3 19 0. 22 5/18 3/3 Chiron QPCR Results #Positive / IU/m #Tested L 21 0. 26 8/18 3/3 26 0. 27 8/16 3/3 28 0. 25 18/18 3/3 35 0. 45 18/18 3/3 *Acrometrix PI. **Combined testing results from multiple sites. 37 0. 36 16/16 3/3 42 1. 15 18/18 3/3 9 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only) NR NR < 3. 3 53. 2 3. 3 23 -28 days 9. 3 27. 0 133. 8 85. 7 910. 0

Clinical Sensitivity: Early Detection of Seroconversion Panel #11008 Day s *ABBOTT PRISM HBs. Ag Clinical Sensitivity: Early Detection of Seroconversion Panel #11008 Day s *ABBOTT PRISM HBs. Ag **Ultrio #Positive / #Tested Chiron QPCR Results #Positive / IU/m. L #Tested 0 0. 32 0/10 0/3 7 0. 17 0/9 0/3 28 0. 18 0/10 0/3 31 0. 28 1/10 0/3 36 0. 25 0/10 2/3 38 0. 22 0/10 1/3 43 0. 27 1/10 1/3 48 0. 23 1/10 1/3 51 0. 24 5/9 3/3 55 0. 30 6/9 3/3 58 0. 29 9/10 3/3 62 0. 39 10/10 3/3 65 0. 61 10/10 3/3 69 10/10 2/2* *Acrometrix 1. 35**Combined testing results from multiple sites PI. 10 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only) NR NR < 3. 3 4. 7 9. 9 5. 4 18 -33 days 38. 8 196. 2 344. 1 193. 4*

Analytical and Clinical Specificities In-house Assay Validation: § 100 random donor samples tested in 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/m. L WHO HBV standard § Potential Interfering Substances Tested • Anticoagulants: ACD, CPD, K 2 EDTA, K 3 EDTA, Na Citrate, Na Heparin • Protein at > 7 g/d. L • Lipid/Triglyceride at > 3000 mg/d. L • Hemoglobin at > 500 mg/d. L • Bilirubin at >20 mg/d. L • Bacteria (>105 CFU/mol) commonly seen in blood related products • Blood-born pathogens: HCV, HTLV, HAV, CMV, EBS, HSV, and HIV-1 11 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only)

Confirming HBV Yield Cases: Chiang Mai University Study (HBs. Ag Negative)* *Nantachit, N. et Confirming HBV Yield Cases: Chiang Mai University Study (HBs. Ag Negative)* *Nantachit, N. et al, in press, 2007. **Later HBs. Ag positive. nd, not determined. 12 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only)

Quantitation of DDL Genotype A-G DDL Genotyp e* Bayer b. DNA 3. 0 Cp/m. Quantitation of DDL Genotype A-G DDL Genotyp e* Bayer b. DNA 3. 0 Cp/m. L A 38367 1997 B Chiron QPCR IU/m. L 9179 1078 NGI Super. Quant Cp/m. L Bayer/Chir NGI/Chir on Cp/IU 28000 4. 2 3. 1 33171 2831 3107 214 4100 10. 7 1. 3 C 32811 1002 2843 250 33000 11. 5 11. 6 D 40197 6429 3328 170 77000 12. 1 23. 1 E 41079 527 2108 141 30000 19. 5 14. 2 F 49087 12332 4955 789 26000 9. 9 5. 2 27003 2278 38000 1. 4 G * Supplied as 37464 Cp/m. L. 30, 000 1558 13 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only)

Summary § A sensitive qualitative and quantitative HBV DNA assay has been developed and Summary § A sensitive qualitative and quantitative HBV DNA assay has been developed and validated in house • LOD at 3. 3 IU/m. L 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? 14 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only)

Acknowledgement § Dennis Madriaga § Baohe Shen § Barney Krebs 15 | Presentation Title Acknowledgement § Dennis Madriaga § Baohe Shen § Barney Krebs 15 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Internal Use Only)