수두 실험실 진단체계 확립 및 수두 바이러스의 분자역학적 분석

초록

Purpose : In this study, we aimed to establish the laboratory diagnostic system for the early detection of varicella zoster virus(VZV), to perform the molecular epidemiological analysis and to provide the basic data for the control and prevention of the chickenpox in Korea. Methods : For the laboratory diagnosis of VZV, we used specimens such as throat swabs, CSF, vesicle swabs collected in the virus transport medium and sera. For the cell culture, we inoculated specimens into A549 or MRC-5 cells and observed cytopatic effects by incubating for 2-3 weeks. For the molecular diagnosis of VZV, we established the nested PCR targeting ORF 38. Using ELISA(Enzygnost Anti-VZV/IgG, IgM, Dade Behring, Germany) kit, we measured the IgM or IgG level of VZV. We also established immunofluorescence method using A549 or MRC-5 cells. For the differentiation of wild type strain, we established the PCR-RFLP analysis of ORF62. Results : By using nested PCR of ORF 38 region, we detected 24 PCR positives among 49 specimens in 2007 and positive strains were all identified as a human herpesvirus 3 strain(HHV—3)and isolated 8 VZV strains in A549 cells. Using the direct immunofluorescence, we confirmed 8 positive specimens. We performed ELISA and obtained 3 VZV IgG positive samples among 14 VZV positive sera. Using PCR-RFLP of ORF62, we confirmed that ail 22 VZV strains except one which were wild type VZV strains. Conclusion : We established laboratory system including nested PCR of ORF 38, cell culture, direct immunofluorescence staining method, and antibody ELISA and the molecular analysis of VZV strains using ORF62 PCR-RFLP. We need to establish the sensitive surveillance and to improve the isolation rate of VZV by the cell culture.

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