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目的 建立简便易行的筛检常见的耐拉米夫定乙肝病毒变异株的方法。方法通过PCR定向点突变 ,构建最常见的耐拉米夫定乙肝病毒变异株 (YMDD变异株 ) ,并建立相应的错配PCR结合限制性片段长度多态性分析 (RFLP)检测此变异株。结果PCR成功引入点突变 ,构建了YMDD变异株的克隆 ,同时错配PCR结合限制性片段长度多态性分析可有效区分YMDD变异株和HBV野毒株。结论采用PCR的方法可诱导定向点突变 ,错配PCR结合限制性片段长度多态性分析可用于筛检临床常见的耐拉米夫定乙肝病毒变异株 (YMDD变异株 ) ,为临床监测提供了一种很好的模式。
Objective To establish a simple and easy method for screening common variants of hepatitis B virus (HBV) resistant to lamivudine. Methods The most common mutant of hepatitis B virus (YMDD) resistant to lamivudine was constructed by PCR directed mutagenesis, and the corresponding mismatch PCR and restriction fragment length polymorphism analysis (RFLP) were used to detect this variant. Results PCR was successfully introduced into the point mutation, and the cloned YMDD mutant was constructed. Meanwhile, mismatch PCR combined with restriction fragment length polymorphism analysis could effectively distinguish YMDD mutant from HBV wild-type strain. Conclusion PCR-directed mutagenesis can be induced. Mismatch PCR combined with restriction fragment length polymorphism (PCR-RFLP) analysis can be used to screen for the clinical variant of hepatitis B virus (YMDD) A good model.