PCR for HBV, HCV and HIV-1 experiences and first results from a routine screening programme 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 by using an automatic sample processor with disposable tips (validated to avoid contamination) taken from blood donations which are serologically negative and free for clinical usage according to federal regulations. In the case of a positive PCR pool result the viraemic donation is identified by two additional 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 additional reverse transcription step, PCR amplifications are performed. PCRs are done for each virus in two genomic regions. Laser-induced detection after PAGE and computer-analysis are used to identify the amplification products. Using this validated methodology routine we have checked 428 896 donations up to the end of August 1996. During this survey we found at least 24 viruses-containing donations which were negative in corresponding serological tests and would have been transfused (2 HBV-, 22 HCV-, 0 HIV-1 -containing donations). It seems possible for large transfusion centres to shorten the diagnostic window periods with our PCR-methodology with acceptable costs (15 DM per donation for all three viruses including logistics, developments and investments).
Schottstedt
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V
Red Cross Blood Transfusion Service North-Rhine Westfalia, Germany.
Tuma
W
W
Bünger
G
G
Lefèvre
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H
D015492:AIDS Serodiagnosis; D001782:Blood Donors; D003365:Costs and Cost Analysis; D005069:Evaluation Studies as Topic; D005188:False Negative Reactions; D005858:Germany; D015658:HIV Infections / Q000175:diagnosis / Q000635:transmission; D015497:HIV-1 / Q000235:genetics* / Q000302:isolation & purification*; D016174:Hepacivirus / Q000235:genetics* / Q000302:isolation & purification*; D006509:Hepatitis B / Q000175:diagnosis / Q000635:transmission; D006515:Hepatitis B virus / Q000235:genetics* / Q000302:isolation & purification*; D006526:Hepatitis C / Q000175:diagnosis / Q000635:transmission; D006801:Humans; D008403:Mass Screening; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods* / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D012698:Serologic Tests; D065227:Transfusion Reaction; D014766:Viremia / Q000175:diagnosis; D014773:Virology / Q000191:economics / Q000379:methods / Q000706:statistics & numerical data