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

2. 2 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Confidential, Internal Use Only) 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. 3 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Confidential, Internal Use Only) 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. 4 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Confidential, Internal Use Only) Analytical Sensitivity and LOD for WHO HBV Standard HBV IU/mL # Positive / # Tested % Positive 2020/20100 1020/20100 520/20100 2.5018/2090 1.2513/2065 0.638/2040 0.313/2015 00/160 HBV (IU/mL) % Positive ED95 3.26 (2.21 – 6.53) IU/mL

5. 5 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Confidential, Internal Use Only) Detection Rate Expected Dose (range), IU/mL Chiron QPCRUltrio*Ultrio dHBV* ED953.3 (2.2–6.5)8.0 (7.1–9.3)6.8 (6.0–7.7) ED500.8 (0.6–1.0)3.3 (3.0–3.8)3.0 (2.7–3.4) Analytical Sensitivity and LOD for WHO HBV Standard *Procleix Ultrio PI, IN0166EN Rev. A

6. 6 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Confidential, Internal Use Only) Analytical Sensitivity for DDL HBV Genotypes HBV Cp/mL Genotype A Genotype B Genotype C Genotype D Genotype E Genotype F Genotype G 1000100 300100 50100 758283100 301007592645083100 109233 8174292 32733 20331790 00000000

7. 7 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Confidential, Internal Use Only) Reproducibility, Linearity, Accuracy, and LLOQ R 2 = 0.995 - 0.999 CV = 0.4% - 1.3% LLOQ = 3.3 IU/mL

8. 8 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Confidential, Internal Use Only) Clinical Sensitivity: Early Detection of Seroconversion Panel #6289 Days *ABBOTT PRISM HBsAg **Ultrio #Positive / #Tested Chiron QPCR Results #Positive / #TestedIU/mL 00.95 0/160/3 NR 20.20 0/170/3 NR 90.56 0/172/3 < 3.3 110.50 2/180/3 NR 160.21 12/173/3 6.2 180.22 16/173/3 14.1 312.15 18/183/3 962.9 364.94 18/183/3 6212.7 15 days *Acrometrix PI. **Combined testing results from multiple sites.

9. 9 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Confidential, Internal Use Only) Clinical Sensitivity: Early Detection of Seroconversion Panel #11006 Days *ABBOTT PRISM HBsAg **Ultrio #Positive / #Tested Chiron QPCR Results #Positive / #TestedIU/mL 00.26 0/170/5 NR 30.16 0/180/5 NR 70.27 0/180/5 NR 100.24 0/160/5 NR 140.24 2/182/3 < 3.3 190.22 5/183/3 53.2 210.26 8/183/3 3.3 260.27 8/163/3 9.3 280.25 18/183/3 27.0 350.45 18/183/3 133.8 370.36 16/163/3 85.7 421.15 18/183/3 910.0 *Acrometrix PI. **Combined testing results from multiple sites. 23-28 days

10. 10 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Confidential, Internal Use Only) Clinical Sensitivity: Early Detection of Seroconversion Panel #11008 Days *ABBOTT PRISM HBsAg **Ultrio #Positive / #Tested Chiron QPCR Results #Positive / #TestedIU/mL 00.32 0/100/3NR 70.17 0/90/3NR 280.18 0/100/3NR 310.28 1/100/3NR 360.25 0/102/3< 3.3 380.22 0/101/3 < 3.3 430.27 1/101/3 < 3.3 480.23 1/101/3 4.7 510.24 5/93/3 9.9 550.30 6/93/3 5.4 580.29 9/103/3 38.8 620.39 10/103/3196.2 650.61 10/103/3 344.1 691.35 10/10 2/2* 193.4* 18-33 days *Acrometrix PI. **Combined testing results from multiple sites

11. 11 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Confidential, Internal Use Only) 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, K 2 EDTA, K 3 EDTA, 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 (>10 5 CFU/mol) commonly seen in blood related products Blood-born pathogens:HCV, HTLV, HAV, CMV, EBS, HSV, and HIV-1

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

13. 13 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Confidential, Internal Use Only) Quantitation of DDL Genotype A-G DDL Genotype* Bayer bDNA3.0 Cp/mL Chiron QPCR IU/mL NGI SuperQuant Cp/mL Bayer/Chiron Cp/IU NGI/Chiron Cp/IU A 38367  1997 9179  1078 28000 4.23.1 B 33171  28313107  214 410010.71.3 C 32811  10022843  250 3300011.511.6 D 40197  64293328  170 7700012.123.1 E 41079  5272108  141 3000019.514.2 F 49087  123324955  789 26000 9.9 5.2 G 37464  155827003  2278 38000 1.4 * Supplied as 30,000 Cp/mL.

14. 14 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Confidential, Internal Use Only) 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. 15 | Presentation Title | Presenter Name | Date | Subject | Confidentiality Level (Strictly Confidential, Confidential, Internal Use Only) Acknowledgement  Dennis Madriaga  Baohe Shen  Barney Krebs