云南和山西无偿献血人群HCV部分基因的核苷酸序列测定及基因分型  

Sequencing and genotyping of the partial gene of HCV among volunteer blood donors in Yunnan and Shanxi province,China

作　　者：王敏[1] 周源[2] 陈帆 王怡仲[4] 许茹[5] 夏文杰[5] 聂咏梅[5] 花文峰[5] 付涌水[5] 

机构地区：[1]中山大学中山医学院生化教研室,广东广州510080 [2]太原市红十字血液中心 [3]昆明血液中心 [4]中山大学生命科学学院 [5]广州血液中心

出　　处：《中国输血杂志》2009年第11期875-879,共5页Chinese Journal of Blood Transfusion

基　　金：国家自然科学基金(项目编号:30872162);广东省自然科学基金(项目编号:06021796);广州市医药卫生重点项目(2006-ZDi-12;2008-ZDi-10)资助课题

摘　　要：目的了解云南和山西2地无偿献血人群丙型肝炎病毒(HCV)的基因型分布及其差异。方法采用荧光定量PCR(Q-PCR),对所收集的203份ELISA检测抗-HCV阳性的无偿献血者标本(云南102份,山西101份)做核酸检测,对核酸阳性的标本做E1基因扩增和测序;E1基因阴性者,进一步做NS5B基因扩增;采用DNASTAR、Bi-oedit和Mega4等软件对E1、NS5B基因进行分析及分子进化研究。结果203份ELISA检测抗-HCV阳性的无偿献血者标本中,云南102份标本经Q-PCR检测出核酸阳性55份,从中扩增出E1基因43份,补做NS5B基因扩增获得阳性结果8份;山西省101份标本经Q-PCR检测出阳性46份,从中扩增出E1基因33份,补做NS5B基因扩增获得阳性结果6份。云南省献血者HCV基因型(亚型)可分为7种:1b[7.84%(4/51)]、2a[13.73%(7/51)]、3a[17.65%(9/51)]、3b[41.18%(21/51)]、6a[7.84%(4/51)]、6n[9.80%(5/51)]和6v[1.96%(1/51)];山西省献血者HCV基因型(亚型)较简单,可分为3种:1b[89.74%(35/39)]、2a[7.69%(3/39)]和3b[2.56%(1/39)]。结论我国云南和山西2地无偿献血人群中的HCV基因型(亚型)的分布存在明显的差异。Objective To investigate the genotypes of HCV among volunteer blood donors in Yunnan and Shanxi provinees of China. Methods Plasma samples were obtained from 102 volunteer blood donors in Yunnan province and 101 volunteer blood donors in Shanxi province. The sampies were all anti-HCV positive. HCV RNA was detected by fluorescent quantitation polymerase chain reaction （Q-PCR）. The positive samples were subjected to E1 gene detection by RT-PCR followed by direct DNA sequencing and Phylogenetic analysis. Samples negative for E1 gene were subjected to NSSB gene detection by RT-PCR followed by direct DNA sequencing and Phylogenetic analysis. Results HCV RNA was detected in 55 samples and genotypes were determined in 51 sampies（43 samples based on E1 gene, and 8 samples based on NSSB gene） from Yunnan province. Among these samples, subtypes lb, 2a, 3a, 3b, 6a, 6n, 6v were detected at frequencies of 7. 84% （4/51）, 13.73% （7/51）, 17.65% （9/ 51 ）, 41.18% （21/51）, 7.84% （4/51） ,9. 80% （5/51） and 1.96% （ 1/51 ）, respectively. HCV RNA was detected in 45 samples and genotypes were determined in 39 samples （33 samples based on E1 gene, and 6 samples based on NSSB gene） from Shanxi province. Among them, subtype 1 b sequences were isolated from 35 samples （89.74%）, 2a sequences were isolated from 3 samples （7.59%）, 3b sequences were isolated from 1 sample （2.56%）. Conclusion Distribution of HCV genotypes among volunteer blood donors was quite different between Yunnan and Shanxi province.

关 键 词：丙型肝炎病毒(HCV) HCV基因型 亚型 基因测序 基因分型 无偿献血者 云南 山西 

分 类 号：R446.6[医药卫生—诊断学] R512.63[医药卫生—临床医学]