Background: Human parvovirus B 19 (B19) is a singlestranded and non-enveloped DNA virus. In the majority of patients infections with B19 are benign and characterised only by malaise and exanthema of the skin. In a small subset of patients B19 infections induce severe complications, including aplastic anaemia, chronic affections of the joints and fetal injuries up to abortion during pregnancy. Since B19 is transmitted through contaminated blood donations, testing for B19 DNA is taken into consideration to further increase virus safety of blood products. Material and Methods: TaqMan technology, a homogeneous and real-time fluorescence PCR approach, has been applied for identification of B19 DNA. Primers were designed that amplify a DNA fragment located within a conserved region of non-structural protein 1 (NS1). A B19 DNA variant showing a 27 base-pair inversion was constructed to be used as internal control. The assay was validated according to the criteria of the Paul- Ehrlich-Institut and European regulatory authorities. Results: The assay had a sensitivity limit of 104 genome equivalents per ml. False-negative results due to problems with amplification were ruled out by the use of an internal control DNA. Reliable quantification of B19 DNA was possible by the use of a standard curve. Conclusions: A real-time PCR procedure for B19 virus DNA detection that is based on the TaqMan technology has been developed and validated. The procedure is applicable for routine testing of single blood donations as well as for analysing large-scale plasma pools.

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© 2002 S. Karger GmbH, Freiburg
2002
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