| 耐拉米夫定乙肝病毒变异株的人工构建及其限制性片段长度多态性分析 |
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| 引用本文： | 孙剑,侯金林,王战会,肖蕾,章廉,骆抗先.耐拉米夫定乙肝病毒变异株的人工构建及其限制性片段长度多态性分析[J].南方医科大学学报,1999,19(6):489-492. |
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| 作者姓名： | 孙剑 侯金林 王战会 肖蕾 章廉 骆抗先 |
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| 作者单位： | 第一军医大学南方医院感染内科广州 51051 5 |
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| 基金项目： | 国家自然科学基金，军队医药卫生重点课题联合资助!(3963028096Z024) |
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| 摘 要： | 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。
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| 关 键 词： | 乙肝病毒 耐药 限制性片段长度多态性 |
| 修稿时间： | 1999年3月8日 |
Construction of HBV lamivudine-resistant mutant and the restriction fragment length polymorphism analysis |
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| Sun Jian,Hou Jinlin,Wang Zhanhui,Xiao Lei,Zhang Lian,Luo Kangxian.Construction of HBV lamivudine-resistant mutant and the restriction fragment length polymorphism analysis[J].Journal of Southern Medical University,1999,19(6):489-492. |
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| Authors: | Sun Jian Hou Jinlin Wang Zhanhui Xiao Lei Zhang Lian Luo Kangxian |
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| Institution: | Sun Jian,Hou Jinlin,Wang Zhanhui,Xiao Lei,Zhang Lian,Luo Kangxian Department of Infectious Diseases,Nanfang Hospital,First Military Medical University,Guangzhou,510515 |
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| Abstract: | Objective To establish an easy method for the detection of HBV lamivudine-resistant mutants Methods HBV YMDD mutant was constructed by site-directed mutagenesis A novel method of mismatched PCR in combination with enzyme digestion was used to distinguish the YMDD mutant from HBV wild type Results The HBV YMDD mutant was proved by sequencing By using the new method of PCR restriction flagment length polymorphism (RFLP), HBV wild type can be easily distinguished from HBV YMDD mutants Conclusion The novel PCR-RFLP can be applied to monitor HBV lamivudine-resistant mutants |
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| Keywords: | Hepatitis B Virus drug-resistance PCR-RFLP |
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