PCR FOR HBV, HCV AND HIV-1 EXPERIENCES AND FIRST RESULTS FROM A ROUTINE SCREENING-PROGRAM IN A LARGE BLOOD-TRANSFUSION SERVICE

We adapted the PCR method to screen up to 3000 blood donations per day for hepatitis B, hepatitis C and HIV-1 virus contamination. Up to 600 aliquots (average 418 donations) are pooled using an automatic sample processor with disposable tips (validated to avoid contamination) tak en from blood donations which are serologically negative and free for clinical usage according to federal regulations. In the case of a posi tive PCR pool result the viraemicdonation is identified by two additio nal PCR pools testing steps with smaller pool sizes. All of the steps are supported by electronic data processing. After virus concentration by ultracentrifugation, and in the case of HCV and HIV-1 an additiona l reverse transcription step, PCR amplifications are performed. PCRs a re done for each virus in two genomic regions. Laser-induced fluoresce nce detection after PAGE and computer-analysis are used to identify th e amplification products. Using this validated methodology routinely w e have checked 428 896 donations up to the end of August 1996. During this survey we found at least 24 virus-containing donations which were negative in corresponding serological tests and would have been trans fused (2 HBV-, 22 HCV-, 0 HIV-1-containing donations). It seems possib le for large transfusion centres to shorten the diagnostic window peri ods with our PCR-methodology with acceptable costs (15 DM per donation for all three viruses including logistics, developments and investmen ts). (C) 1998 The International Association of Biological Standardizat ion.