3’碱基特异性聚合酶链反应快速筛查基因点突变

3' Base specific polymerase chain reaction for rapid screening assay for genetic point mutation

目的建立并评价检测乙型肝炎病毒(HBV)基因前C区1896位点突变的3’碱基特异性聚合酶链反应(3’BS-PCR)。方法以3’BS-PCR法结合PCR产物直接测序法测定72例抗-HBe阳性慢性乙型肝炎患者HBV前C1896位点突变。结果在61℃最佳退火温度条件下,16例慢性乙型肝炎患者3’BS-PCR测得的点突变结果与测序法结果完全相符;利用3’BS-PCR法检测72例抗-HBe阳性慢性乙型肝炎患者,前C1896位总突变率为34.72%,其中单纯突变感染率为19.44%,混合株感染率为15.28%。结论3’BS-PCR方法操作简单经济,提供了有应用价值的大规模筛检DNA点突变的方法。

Objective To evaluate the 3'base specific polymerase chain reaction(3'BS-PCR) screening method for detecting pre core (pre-c) 1?896 point mutations of hepatitis B virus(HBV).Methods Pre-c 1?896 point mutations of HBV were detected in 72 cases of patients with chronic hepatitis B infection by 3'BS-PCR, as well as direct sequencing of PCR product.Results In this study when the anneal temprature was 61 ℃, the 3'BS-PCR results of pre-c point mutation in 16 cases were completely conformed to that of direct sequencing of PCR product; The rate of pre-c point mutation in 72 case of anti-HBe positive patients was 34.72% when using 3'BS-PCR, within which the pure mutation rate and overall mutation rate were 19.44% and 15.28% respectively.Conclusion 3'BS-PCR, a simple and economical method, could support us with an alternatively valuable assay for screening DNA point mutations from large number samples.