Development of the Enzyme Immunoassay for the Detection of Anti-HSV-2 Antibody with HSV-2 Specific Monoclonal Antibody / 감염

ABSTRACT

BACKGROUND: The serological diagnosis of herpes simplex virus type 2 (HSV-2) infection has pitfalls, in that most of the antibodies against HSV-2 cross-react with HSV-1 and the prevalence of HSV-1 infection is high, especially in Korea. In this study, we tried to establish the serological diagnostic method, which could detect and measure the specific antibodies against HSV- 2 by competitive immunofluorescent staining method as well as competitive ELISA based on the specific monoclonal antibody, MH2-7. METHODS: Immunofluorescent staining and western blot analysis were used to characterize the antigens recognized by MH2-7. Competitive immunofluorescent staining (IF), competitive enzyme immunoassay (ELISA), and western blot analysis were used to detect specific antibodies against HSV-2 in patients' sera. RESULTS: In western blot analysis, the sera from two of six patients clinically diagnosed as genital herpes showed characteristic band patterns, which have been known to be compatible with HSV-2 infection. In competitive immunofluorescent staining, only the sera from the two patients clinically diagnosed as genital herpes and with characteristic band pattern showed competition with MH2-7 monoclonal antibody. The dilution range of the serum showing specific competition was between 110 and 180. Competitive ELISA was also performed and evaluated as the diagnostic efficacy as ELISA has been known to be advantageous over IF staining in mass screening. The result showed linear dose-response relationship for the patient's sera in inhibition of the reactivity of MH2-7. CONCLUSION: We suggest that the competitive immunofluorescent staining method and competitive ELISA based on the specific monoclonal antibody MH2-7 is a simple, accurate, and precise method, which can be used in serological diagnosis of HSV-2 infection.