| 三种检测乙型肝炎病毒YMDD自然变异方法的比较 |
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| 引用本文： | 闵晓春,赵书民,杨德刚.三种检测乙型肝炎病毒YMDD自然变异方法的比较[J].实用肝脏病杂志,2010,13(1):16-18,25. |
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| 作者姓名： | 闵晓春 赵书民 杨德刚 |
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| 作者单位： | 1. 上海市嘉定区中心医院感染病科,201800
2. 上海第二军医大学附属长征医院感染病科 |
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| 摘 要： | 目的筛选一种特异性及灵敏性较高的检测乙型肝炎病毒YMDD自然变异的方法。方法通过分子克隆的方法,构建YMDD、YVDD及YIDD三种质粒;通过检测不同比例混合的质粒,比较PCR产物直接测序法、单探针特异性荧光PCR技术及引物特异性荧光PCR技术三种检测方法。结果经酶切鉴定及直接测序鉴定,显示三种质粒构建成功;当待测模板占总模板的比例大于20%时,直接测序法才能准确检测;当突变株占总毒株的比例占50/51时,单探针特异性荧光PCR法才能判定该样品为YMDD变异型;当突变株占总毒株的比例高于1/11时,引物特异性荧光PCR技术非特异结合的影响可忽略,具有很好的特异性,同时具有较高的灵敏性,检测突变株占总毒株的最低比例可达到1/11。结论引物特异性荧光PCR技术特异性及灵敏性较高,适合用于乙型肝炎病毒YMDD自然突变株的检测。
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| 关 键 词： | 乙型肝炎病毒 YMDD自然变异 PCR |
The comparison of three assays on the spontaneous YMDD mutation in Hepatitis B Virus |
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| MIN Xiaochun,ZHAo Shumin,YANG Degang.The comparison of three assays on the spontaneous YMDD mutation in Hepatitis B Virus[J].Journal of Clinical Hepatology,2010,13(1):16-18,25. |
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| Authors: | MIN Xiaochun ZHAo Shumin YANG Degang |
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| Institution: | MIN Xiao-chun,ZHAO Shumin,YANG Degang. Department of Infectious Disease,Jiading District Central Hospital,Shanghai 201800,China |
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| Abstract: | Objective To sieve a assay of higher specificity and sensitivity on the spontaneous YMDD mu-tation in hepatitis B virus. Method Plasmids of YMDD,YVDD,YIDD were constructed with molecular methods. The sensitivity and specificity of the three assays (direct sequencing of PCR products,single probe-specific fluo-rescent PCR and primer-specific fluorescent PCR)were compared by detecting coexisting strains. Result Plasmids of YMDD,YVDD,YIDD which were constructed successfully were identified by restriction enzyme... |
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| Keywords: | Hepatitis B virus YMDD spontaneous mutation PCR |
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