3’碱基特异性聚合酶链反应快速筛查基因点突变

汲振余; 赵翠林; 陈茜; 李程; 朱金凤

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

摘要

摘要:
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。

Abstract:
Objective To evaluate the 3'base specific polymerase chain reaction(3'BS-PCR)screening method for detecting precore(pre-c)1896 point mutations of hepatitis B virus(HBV).
Methods Pre-c 1896 point mutations of HBV were detected in 72 cases of patients with chronic hepatitis Binfection 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.

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