山西省吸毒人员丙型肝炎病毒的基因分型和序列分析

Study on Genotype and Sequencing of Hepatitis C Virus (HCV) among Drug Addict Population in Shanxi

目的 探讨山西省吸毒人群中HCV基因型分布现状和基因特征。方法 采集太原强制戒毒所576份吸毒人员血清标本,用ELISA法进行抗HCV的检测;检出抗HCV阳性的血清进一步用型特异性引物RT-巢式PCR法进行基因分型,选择1b型和2a型各一株进行PCR产物直接测序。结果 576份吸毒人员的抗HCV检出率为7.5％其中本省的抗HCV检出率为5.7％,外省为27.7％,外省吸毒者的抗HCV检出率非常显著高于山西省吸毒者(X^2=30.32,P<0.01)。山西省吸毒人群的HCV基因型以1b型为主(78.9%),其次为2a型(15.8%)和1b/2a混合型(5.3％);未检出1a型、2b型和3a型,但山西省分型阳性者中口吸者居多(84.2％)。山西吸毒者HCV 1b型分离株在其核心区的144个核苷酸序列中,与日本、河北、上海、湖南及邻省的同源性都在94％以上,与湖南株的同源性最好(97％);而推导的氨基酸序列与日本的差异最大(7％)。山西吸毒者HCV 2a型分离株在其核心区的174个cD-NA核苷酸序列和推导的氨基酸序列中与日本的同源性分别为94％和96％,但氨基酸的变异小于核苷酸。值得注意的是,nt 522和 nt 564(核心区第209和223位)的变异是所有中国1b型株的共同特征。结论 山西省吸毒人群中HCV基因型以1b为主。在同一亚型中,山西吸毒者的HCV分离株在核心区部分序列与日本及国内其他地区的分?

Objective To explore the situation of HCV genotypes distribution and gene characteristic in population of drug users in Shanxi.Method 576 sera samples of drug user were collected, and HCV antibodies were detected by ELISA for those samples. The genotyping of HCV was conducted on anti - HCV positive samples using type specific primers deduced from the core region of HCV by RT- nest PCR, then direct sequencing was preformated for two PCR products (genotype Ib and 2a) .Result The anti -HCV positive rate of drug users was 7.5% .The drug users from other cities had higher anti - HCV positive rate (27.7% ) than that of drug users (5.7%) from Shanxi (X2 = 30.32, P < 0.01) .1b (78.9%) is the predominant genotype of HCV in drug users in Shanxi.The next is 2a (15.8%) and lb/2a (5.3%) .No genotype of la, 2b, and 3a were detected.The nucleotide homogenous rate of 1b for Shanxi strain comparing with HC - J4/83, strains of Herbei, Hunan, Shanghai is all higher than 94% , and more close to Hunan. But the derived ammo acid has largest different from HC - J4/83 (7 % ) . The homogenous of 2a for Shanxi strain with HC -J6 is 94% (96% for amino acid) .The homogenous of derived amino acid sequence was better than nucleotide sequence in corresponding region among all strains.The mutation at nt 522 and nt 564 (core nt 209 and nt 223) is the common characteristic of all genotype Ib strains in China in this study. Conclusion The predominant genotype of HCV in drug users of Shanxi is 1b.The variation in core region of HCV from drug user in Shanxi is existed comparing to strains of Japan and other regions of China, without significance.