1. SoGAT, Berne 2006BTS SRC Berne Ltd. Evaluation of the Procleix TIGRIS System at the Blood Transfusion Service Berne Ltd. Dr. Martin Stolz

2. SoGAT, Berne 2006BTS SRC Berne Ltd. NAT testing in progress HCV-PCRsince 01 ‘1999 HIV-1 PCRsince 03‘ 2002 increased blood safety automized test procedures

3. SoGAT, Berne 2006BTS SRC Berne Ltd. Procleix TIGRIS System fully automated NAT screening system combined detection of 3 viruses: HIV-1, HCV and HBV extraction/amplification/detection in one tube extraction based upon magnetic bead technology amplification based upon the TMA technology using RNA as amplified targed detection by chemiluminescence specific discrimination assays for virus identification

4. SoGAT, Berne 2006BTS SRC Berne Ltd. Design of the evalution study Assay validation: – sensitivity, specificity, robustness & cross-contamination Test performance under routine conditions – routine samples – pools of 8 or individual donations – pooling with Tecan pipetting robots – analysis with Procleix TIGRIS system

5. SoGAT, Berne 2006BTS SRC Berne Ltd. Sensitivity HIV-1:24.9 IU/ml (95 % C.I.: 20.2 – 33.3 IU/ml) ca. 13.9 copies/ml HCV: 3.7 IU/ml (95 % C.I.: 3.1 – 4.8 IU/ml) ca. 23.3 copies/ml HBV:7.6 IU/ml (95 % C.I.: 6.0 – 11.1 IU/ml) ca. 41.0 copies/ml

6. SoGAT, Berne 2006BTS SRC Berne Ltd. Robustness and reproducibility Procleix TIGRIS system: excellent robustness against cross-contamination reproducibility: intra-, inter-assay precision and between laboratories confirmed reliable test: 3 x 95% positive cut off value (HIV-1: 75 IU/ml, HCV: 11 IU/ml, HBV: 23 IU/ml) high robustness against inhibiting substances i.e. 100 I.U. heparin per sample no interference with lipaemic or haemolytic samples as a frequent donation related factor

7. SoGAT, Berne 2006BTS SRC Berne Ltd. Specificity – System Evaluation *) positive sample was a HBV window period donation

8. SoGAT, Berne 2006BTS SRC Berne Ltd. Summary very good test sensitivity, robustness and reliability clear discrimination between reactive and non reactive samples low level of invalid samples elevated level of „false“ positives with pools of 8 observed pool resolving requires more time and large sample quantities fully automated NAT screening system test and software easy to handle high throughput: single donation testing feasable very high degree of automatisation, but demanding hardware and little flexibility

9. SoGAT, Berne 2006BTS SRC Berne Ltd. Acknowledgments Christiane Schmidseder Rita Hubschmid Roger Hellmüller Dr. Christoph Niederhauser Dr. Philippe Schneider (BTS Lausanne) Chiron Corp.

10. SoGAT, Berne 2006BTS SRC Berne Ltd. Window period HBV donation identified by single donation testing donation already transfused at time of testing recipient developed a HBV viraemie 3 weeks post transfusion viral load of the initial donation not available, but… initial HBV titer must have been quite low: no longer detectable in a dilution of 1:8 and 1:24 101'621 IU/ml in 2 nd donation 35 days later