Failure to detect hepatitis A virus RNA in factor VIII concentrates using the polymerase chain reaction. Results of a preliminary study.

The transmission of hepatitis A virus (HAV) infection to recipients of some batches of Factor VIII has recently been reported. The polymerase chain reaction (PCR) was used for the detection of HAV RNA in factor VIII concentrates. Primer sequences used were derived from a consensus of published sequences in the 5' non coding region; a nested PCR was used to increase sensitivity and specificity and the resulting fragment was 151 base pairs in length. The PCR was initially validated in clinical samples and only IgM anti-HAV positive patient samples and a sample of liver tissue from a patient who required liver transplantation for fulminant hepatitis A were HAV PCR positive. Other samples tested included those that were IgG anti-HAV positive; these were found to be PCR negative. In an investigation of coagulation factor VIII concentrates by HAV PCR, 40 batches of solvent/detergent-treated high-purity concentrate from four different manufacturers, including one batch of factor VIII possibly implicated in HAV transmission, and a further 3 batches of monoclonal antibody purified factor VIII were all HAV PCR negative. Gel chromatography material, before and after use in factor VIII purification, and eluates from this material were also negative for HAV RNA. Our preliminary results therefore suggest that either the contamination of factor VIII concentrates by HAV RNA is an extremely rare event or that the PCR is insufficiently sensitive to detect an infective HAV dose since each batch of factor VIII concentrate would have been derived from a plasma pool consisting of 10,000 donations, or more and the resulting concentration of virus may be 10(2) or less.