PCR产物直接测序检测乙型肝炎病毒耐药变异位点  

Clinical application of DNA sequencing for detecting point mutations in hepatitis B virus associated with drug resistance

作　　者：孔令和[1] 高素香[1] 柯云霞[1] 杨剑[1] 雷桂萍[1] 史国香[1] 

机构地区：[1]江西省九江市第三医院,江西九江332000

出　　处：《南方医科大学学报》2012年第2期277-279,共3页Journal of Southern Medical University

摘　　要：目的研究采用PCR产物直接测序技术在乙型肝炎病毒(HBV)耐药变异位点检测的特异性和实用性。方法 166例经过核苷类似物治疗至少2年以上的慢性乙肝患者,提取血清HBVDNA,采用巢式PCR扩增,将所得的PCR产物进行琼脂糖电泳检测,阳性PCR产物直接测序。对测序成功的样本进行耐药变异分析。结果 166例中有120例HBV DNA扩增电泳产物阳性,测序确定基因耐药变异位点有40例(24.1%,40/166);其具体分布为:L180M和M204V/I均变异17例(42.5%,17/40)、M204V/I变异lO例(25%,10/40)、N236T变异8例(20%,8/40)、L180M变异3例(7.5%,3/40)、A181V/T变异1例(2.5%,1/40)、A181V/T和N236T变异1例(2.5%,1/40)。120例患者测序确定基因耐药未测出变异位点有80例(66.66%,60/120)例。结论本院核苷类似物治疗2年以上患者,控制病毒疗效并不佳,有24%患者出现基因耐药。PCR产物直接测序法是目前检测HBV耐药变异的一种较好的方法,可用于专科医院临床检测。Objective To assess the specificity and applicability of direct PCR sequencing in the detection of point mutations in hepatitis B virus（HBV） associated with drug resistance.Methods Serum samples were obtained from 120 patients with hepatitis B treated with nucleoside analogus for at least 2 years to detect the point mutations in HBV genome in association with drug resistance using nested PCR and direct DNA sequencing.Results Forty out of the 120 patients were found to have one or two point mutations associated with drug resistance,including 17 with L180M and M204V/I mutations（42.5%）,10 with M204V/I mutation（25%）,8 with N236T mutation（20%）,3 with L180M mutation（7.5%）,and 1 with both A181V/T and N236T mutations（2.5%）,and 1 with A181V/T mutation（2.5%）.Conclusion DNA sequencing is a good method to detect all known point mutations associated with HBV drug resistance.

关 键 词：乙型肝炎病毒 耐药突变 测序检测 

分 类 号：R440[医药卫生—诊断学]