국내 유행 HCV를 사용한 Replicon cell 제작 및 유전자 변이 분석

초록

Purpose : In order to survey the prevalence of HCV in Korea, we performed analysis of HCV genotypes among chronic hepatitis C patients. Full genome sequences are obtained from prevalent strains such as HCV genotype lb and 2c for infectious and replicable replicon construction. Methods : Serum samples were collected from chronic hepatitis C patients in Korea. Viral RNA was purified from sera and subsequently cDNA synthesis was performed to determine HCV genotype by using INNO-LiPA HCVⅡ (INNOGENETICS，Ghent, Belgiu). Six and 7 amplicons covering HCV full genome were obtained from HCV lb and 2a，respectively. After cloning the target amplicons，the sequences were analyzed by a multiple-alignment algorithm(Clustal Method) in the MegAlign package(Windows version 3.12e; DNASTAR，Madison，WI，USA). Results : The prevalent HCV genotypes were genotype lb and 2a in chronic hepatitis C patients. About 9.3 kbp nucleotide sequence was obtained from HCV genotype lb showing 85% homology comparing with HCV lb strain ND24(AF207765). In the case of HCV genotype 2a，88.9-92.8% homology was observed comparing with HCV reference strains as AB047639，AF169002，AY746460 and D00944. The vector system for infectious and replicable replicon of HCV was constructed. Conclusions : The most prevalent HCV genotype was genotype lb in Korea，whose whole sequence showed 85% homology comparing with AF207765. In the case of HCV genotype 2a，88.9-92.8% homology was observed comparing with HCV reference strains.

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