双探针实时荧光定量PCR法检测对拉米夫定耐药的乙型肝炎患者HBV变异  

Application of dual-probe real-time PCR assay for detection of lamivudine-resistance hepatitis B virus

作　　者：凤敏华 吴婴 黄荣辉 

机构地区：[1]上海市奉贤中心医院核医学科,上海201400

出　　处：《临床检验杂志》2010年第3期175-176,共2页Chinese Journal of Clinical Laboratory Science

摘　　要：目的研究双探针实时荧光定量PCR方法检测对拉米夫定耐药的乙肝患者血清中HBV病毒变异的可行性。方法用双探针MGB实时荧光定量PCR法检测拉米夫定治疗后的乙型肝炎患者血清,确定HBV YMDD变异类型,与测序法进行比对。结果68例标本中本法测出27例YMDD野生株,20例Y IDD变异株,11例YVDD变异株,与直接测序结果一致。另有10例标本测得为混合株,经克隆后测序证实,混合株中存在优势株。直接测序只能检测出优势株。结论双探针MGB实时荧光定量PCR法可以快速、准确地测出HBV DNA混合变异毒株。Objective To investigate the feasibility of dual-probe real-time PCR assay for detection of lamivudine-resistance hepatitis B virus.Methods MGB probes with different fluorescences were designed and synthesized.The serum samples of hepatitis B patients treated with lamivudine were detected by real-time PCR and sequencing,and the results were compared.Results In total 68 serum samples,27 samples are YMDD,20 YIDD and 11 YVDD,which was consistent with that of sequencing（85.3%）.10 samples were detected to be mixed population.Further sequencing assay after cloning demonstrated that the direct sequencing method could only give sequence of major virus in mixed population.Conclusions MGB dual-probe real-time PCR assay can quickly and accurately detect HBV YMDD mutations especially in mixed population.

关 键 词：实时荧光定量PCR 乙型肝炎病毒 拉米夫定 YMDD变异 

分 类 号：R512.6[医药卫生—内科学]