突变特异PCR检测YMDD突变方法的建立及应用  

Construction and application of mutation specific PCR method for detecting YMDD mutation of HBV

作　　者：崔荣辉[1] 戴二黑[1] 陈翠英[1] 宋亚军[2] 杨瑞馥[2] 

机构地区：[1]河北省石家庄市第五医院,050021 [2]军事医学科学院微生物流行病研究所

出　　处：《河北医药》2008年第9期1282-1283,共2页Hebei Medical Journal

基　　金：河北省医学适用技术跟踪项目(编号:32099);石家庄市科技攻关项目(编号:00200716)

摘　　要：目的建立检测YMDD突变的方法并了解乙型肝炎后肝硬化患者采用拉米夫定治疗后YMDD突变的状况。方法采用突变特异PCR技术研究YMDD突变。结果采用突变特异PCR技术和PCR-RFLP方法同时对30份HBV DNA阳性血清标本进行检测,2种方法的YMDD突变检出率基本相符。120例肝硬化患者治疗前无1例检测到YMDD突变,拉米夫定治疗组患者治疗6、12和24个月YMDD突变率分别为0(0/60)、8.5%(5/59)和17.9%(10/56),而对照组患者没有1例发生YMDD突变。结论突变特异PCR技术检测YMDD突变简便特异。YMDD突变的发生率随着拉米夫定用药时间的延长而增高。Objective To establish mutation specific polymerase chain reaction（msPCR） method for detecting YMDD mutation of HBV, and study the YMDD mutation rate of liver cirrhosis after lamivudine treatment. Methods According to YMDD mutation characteristics, the primers were designed and synthesized and msPCR was established, Results Thirty HBV DNA positive sera were analyzed simultaneously by msPCR and PCR-ELISA methods, and the results of two methods were basically consistent, No YMDD mutation was detected in 120 cases of liver cirrhosis before lamivudine treatment. During 6, 12 and 24 months of treatment, YMDD mutation rate was 0（0/60）, 8.5 % （5/59） and 17.9% （10/56） respectively in lamivudine treatment group, but no YMDD mutation was identified in control group. Conclusion The msPCR method for detecting YMDD mutation was simple and specific. Mutation rate of YMDD becomes higher with prolonging of lamivudine treatment.

关 键 词：肝炎病毒 乙型 肝硬化 拉米夫定 突变 聚合酶链反应 

分 类 号：R512.62[医药卫生—内科学] R446.5[医药卫生—临床医学]