DEVELOPMENT OF A NEW CONCENTRATION METHOD OF VIRUS USING POLY-L-LYSINE-COATED MAGNETIC BEADS: APPLICATION TO THE DETECTION OF OCCULT HBV

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  • ポリ‐Ｌ‐リジンコート磁性粒子を用いたウイルスの新規濃縮法の開発‐ｏｃｃｕｌｔ　ｈｅｐａｔｉｔｉｓ　Ｂ検出への応用‐

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Background: The risk of post-transfusion hepatitis B virus (HBV) infection has been reduced following the implementation of HBV nucleic acid amplification technology (NAT). However, the problem of HBV-DNA-positive and HBsAg-negative occult HBV infections remains to be solved. Here, in an attempt to resolve such problems, we report the development of a convenient method to concentrate HBV and HBs-antigen (HBsAg) in order to improve detection sensitivity.<br> Objects: Seventy-eight anti-HBc-positive (≥25: HI), HBsAg-negative and 20-pool NAT negative donations were examined.<br> Methods: Virus concentration is achieved by the enhancement of the agglutination of viruses using poly-L-lysine-coated magnetic beads in the presence of a bivalent metal. HBsAg and HBV-DNA levels were concentrated up-to 20-fold and 8-fold respectively.<br> HBsAg and anti-HBc were tested by enzyme immunoassay (EIA) (AxSYM: Abbott) and hemagglutination inhibition test (HI: Japanese Red Cross) respectively.<br> Results: Of 78 anti-HBc-positive and HBsAg-negative donors, 16 were HBV-DNA-positive by individual NAT and 62 were HBV-DNA-negative by individual NAT. Twenty five of 62 HBV-DNA-negative by individual NAT became HBV-DNA-positive by HBV concentration. Twenty nine of 78 donations which were HBsAg-negative by conventional EIA became HBsAg-positive by HBsAg concentration.<br> Conclusion: Our new method of concentrating HBV and HBsAg increased the sensitivities of EIA and HBV NAT and enabled us to detect HBV-DNA from individual-NAT-negative samples and HBsAg from conventional-EIA-negative samples.<br>

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