HBV BCP区1762/1764和1896突变位点检测新技术研究  

The Study on Mutation Locus Detection Technology of the HBV BCP Double Mutations with 1762 /1764 and Precore Mutation with 1896

作　　者：王泽 唐景峰[2] 

机构地区：[1]阳泉市第三人民医院,山西阳泉045000 [2]武汉大学生命科学学院/病毒学国家重点实验室,湖北武汉430072

出　　处：《山西职工医学院学报》2014年第4期8-13,共6页Journal of Shanxi Medical College for Continuing Education

摘　　要：目的:利用错配扩增和荧光PCR原理,建立一种针对HBV BCP区1762/1764和前C区1896突变位点的新型检测方法——错配扩增突变分析PCR法(MAMA-PCR),并对该方法进行临床评价。方法:a)MAMAPCR突变检测方法性能评估:以野生和突变性阳性参考品作为模板检测3次,分析该方法稳定性;将突变型和野生型质控品梯度比例混合,分析该方法检测混合感染的检测效率。b)测序法、商品化试剂盒与本方法同时检测样本并对MAMA-PCR方法做出评价。结果:MAMA-PCR 3次检测重复性良好,CV值均小于5%,且可稳定检出含1%突变含量的混合样本。132例HBV样本中,1762/1764双突变位点测序法检出67例,ARMS-PCR试剂盒检出69例,MAMA-PCR检出69例,突变检出率分别为50.8%、52.3%和52.3%;1896突变测序法检出39例,ARMSPCR试剂盒检出41例,MAMA-PCR检出42例,突变检出率分别为29.5%、31.1%和31.8%。结论:MAMA-PCR检测方法性能稳定,突变检出率高于测序法,与商用试剂盒基本一致,可用于临床乙肝患者BCP区1762/1764双突变和前C区1896突变位点的快速检测。Objective：To establish a new method（ MAMA-PCR）for the detection of mutations at 1762/1764 and 1896 in the BCP region and precore region of HBV using mismatch amplification and Fluorescent PCR and to evaluate this method with clinical samples. Method：a）Evaluate the parameters of the MAMA-PCR mutation detection method：to analyze the stability of reagents by reactions with three repeats using the wild-type and mutant reference as a template;to test the detection efficiency of the HBV hybrid infection using mutant and wild-type references with different mixing ratio. b）Compare the MAMA-PCR method with sequencing and commercialized mutation detection kits and evaluate the MAMA-PCR method. Results：Three tests of MAMA-PCR had good repeatability with a CV value of less than 5% and HBV infection samples which contained 1% mutation could be stably detected. In 132 HBV samples,sequencing method identified 67 with 1762/1764 mutations,mutation detection kit identified 69,and MAMA-PCR identified 69. The detection rates were 50. 8%,52. 3%,and 52. 3%,respectively. Sequencing method identified 39 samples with 1896 mutation,mutation detection kit identified 41,and MAMA-PCR identified 42. The detection rates were 29. 5%, 31. 1%,and 31. 8%,respectively. Conclusions：The MAMA-PCR method is very stable. The mutation detection rate of MAMA-PCR is almost the same as commercialized kits,and is higher than sequencing. This new technology（ MAMA-PCR）can be used in the fast detection of HBV BCP 1762/1764 double mutations and precore 1896 mutation in clinical samples.

关 键 词：HBV BCP区双突变 前C区突变 MAMA—PCR 

分 类 号：R446.7[医药卫生—诊断学]