VALIDATION OF A PCR ASSAY SYSTEM TO SCREEN PLASMA FOR HBV, HCV, AND HIV-1

We describe a PCR system for the screening of plasma for fractionation , which detects nucleic acids of hepatitis B and C viruses (HBV and HC V) and of human immunodeficiency virus type 1 (HIV-1). Pilot pools of different sizes (12, 120 and 1,200 donations) are generated, and a cas cade testing is employed in order to identify PCR-positive samples. A virus concentration step is included in the initial screening of the l arge pools. The comprehensive efforts in validating the PCR methodolog y as well as the whole system are described; the data demonstrate that the system is well-suited for high throughput testing and that the me thod is sensitive, robust and reproducible. Donations with virus loads greater than or equal to 1.2 x 10(3) genome equivalents/ml (GE/ml) fo r HBV, greater than or equal to 8.4 x 10(3) GE/ml for HCV, and greater than or equal to 4.9 x 10(4) GE/ml for HIV-1 are eliminated with 95% confidence. Therefore, this system will contribute to the virus safety net for the final plasma product.