反义抑制PCR检测乙肝病毒G1896A变异株方法的研究

Method Research of the Antisense Inhibit PCR Detection of Hepatitis B Virus G1896A Mutant

目的:研究和评估反义抑制PCR检测乙型肝炎病毒(HBV)基因前C区1896位点变异株的方法。方法:根据HBV DNA前C区1896位点碱基的突变,设计一系列引物,其中之一是针对该HBV DNA1896位点野生型碱基的反义引物,PCR扩增时,通过对1896位点野生型碱基序列的竞争性抑制,而阻滞野生株DNA的扩增,从而仅对HBV DN A1896位变异的碱基序列扩增,特异性检测变异株。为了评估该方法的特异性,使用限制性内切酶方法对检测结果进行对比。结果:该方法检测HBV G1896A变异株具有特异性,而且检测的最低值可达5×104IU/ml。对慢性乙肝病例82份,检出突变型样本21例,阳性率25.61%,与限制性内切酶方法检测比较,经统计学处理,χ2检验评价两者无显著意义。结论:该法在对HBV临床标本1896位点变异株检测中特异和灵敏,是一种有效的检测方法。

Objective：Research and evaluation the Method of the antisense inhibit PCR detection of hepatitis B virus G1896A mutant.Methods：According to the HBV D N A former area C 1896 sites mutations,design series primers,One is for the HBV D N A wild type of antisense inhibit primer,PCR,Through the wild type of 1896 loci base sequences of competitive inhibition and block the amplification of wild plant DNA.Thus only A1896 bits of HBV DNA base sequences amplification,specificity of detection variants.In order to evaluate the specificity of the method,the use of Restriction enzymes method to test results were compared.Results：This method of detection HBV G1896A mutant with specificity,The lowest test can reach 5 x104 IU/ml.The Cases of 82 of chronic hepatitis b is detected,mutation rate of 21 cases of samples,25.61%,and by detecting comparison Restriction enzymes method,statistical treatment,X-square test evaluation both no significant.Conclusion：This law in clinical specimens of HBV mutant detection in 1896 site is specific and sensitive,it is a kind of effective method.