HBV耐药基因的实验研究

Study on the experiment of drug resistance gene in Hepatitis B Virus

目的:利用多色荧光PCR技术,建立快速检测HBV耐药基因的方法,并与基因测序法进行对照研究。方法:建立2个反应体系,每个反应体系包含四个耐药位点。对新建的多色荧光PCR法进行灵敏度和特异性分析,并对42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1×103copies/ml。42例样本中检测到YVDD 3例(占7.14%),YIDD 1例(占2.38%);1896变异7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,较基因测序法方便、快捷。

Objective：To set up a rapid method for detection of drug resistance mutation in HBV based on multicolour real time PCR. To compare the method with gene sequencing. Methods： Two reaction systems were established with each containing four resistance loci. On the new multicolor real time PCR method, the sensitivity and specificity were analysed, and the drug re- sistant mutation of blood serum collected from 30 patients was detected. Results： A system of multicolor fluorescence PCR for de- tecting drug resistance in HBV was constructed with better specificity and the sensitivity of 1 × 10^3 copies/ml. Among the 42 samples, there were 3 cases of YVDD（7.14% ）, 1 case of YIDD（2.38% ） and there were 7 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 in the clinical hospitals, which would be more rapid and convenient than gene se- quencing.