多色荧光PCR法检测HBV耐药位点突变  

Development of multicolour real time PCR for detection of drug resistance mutation in hepatitis B virus

作　　者：陈兆军[1] 张腊红[1] 朱明利[2] 刘霞[1] 潘峰[1] 徐立群[1] 赵晓飞[1] 林冰[1] 

机构地区：[1]杭州师范大学附属医院检验科,310015 [2]杭州市第六人民医院开放实验室

出　　处：《中华实验和临床病毒学杂志》2010年第6期473-475,共3页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的 利用多色荧光PCR技术,建立一套快速检测HBV耐药位点突变的方法 ,并对临床收集的服药患者血清进行耐药位点检测.方法 建立2个反应体系,每个反应体系包含4个耐药位点.对新建的多色荧光PCR法进行灵敏度和特异性分析,并对30例临床收集的服药患者血清进行耐药检测.结果 构建了多色荧光PCR检测HBV耐药位点的体系,其特异性较好,分析灵敏度可达1×103拷贝/ml.30例样本中检测到YVDD 2例（占6.67%）,YIDD 1例（占3.33%）;1896位点变异5例,占总数的16.67%.其他位点未检测到耐药突变.结论 所构建的多色荧光PCR检测体系为临床医院提供快速、简便的HBV耐药位点突变检测手段.HBV基因前C区的1896位点变异率较高.Objective To set up a rapid method for detection of drug resistance mutation in HBV,based on multicolour real time PCR. To detect the mutation of blood serum, which were collected from patients. Method To establish two reaction systems, each reaction system contains four resistance loci. On the new multicolor real time PCR method, the sensitivity and specificity were analysed, and detecting the drug resistant mutation of blood serum collected from 30 cases patients. Results Construction of a multicolor fluorescence PCR for detection drug resistance in HBV was constructed, better specificity,sensitivity analysis of up to 1 × 103 copies/ml. Sample of 30 cases were detected, there were 2 YVDD （6.67%）, 1 YIDD （3.33%）, and there were 5 cases of 1896 variation, accounting for 16. 67%. Other sites were not detected mutations. Conclusion The multicolor real time PCR detection system could be used for rapid and simple analysis of drug resistance for the clinical hospital. The 1986 mutation in HBV pre-C region are relatively high.

关 键 词：聚合酶链反应 肝炎病毒 乙型 药物耐受 病毒 突变 

分 类 号：R440[医药卫生—诊断学]