目的 应用分子信标探针检测结核杆菌耐异烟肼kat G463condon点突变,并与测序结果比较以验证该检测方法.方法 运用软件对Beacon designer设计,katG基因包含463condon的分子信标,建立其扩增体系及分子信标芯片检测方法,对扩增产物测序并作比较.结果 通过CDC相机观测到结核标准株及耐异烟肼PCR产物与分子信标杂交后荧光信号区别明显;16株耐异烟肼组与10株H37RV标准株对照组荧光信号强度比较,耐异烟肼组463condon突变检出率为37%,分子信标检测方法与测序法符合率达93%.结论 分子信标技术是一种具有高灵敏核酸点突变检测技术;分子信标芯片检测方法与测序法符合率较好.%Objective To detect the mutation site of codon 463 in katG gene of INH resistant MTB by molecular beacon probe, and verify the detecting method by comparing with gene sequencing. Methods The software Beacon designer,was used to design codon 463 molecular beacon probe and the amplification system,and then the method was compared with gene sequencing. Results The difference between PCR products from standard strain and INH resistant one is obvious in detecting the fluores-cent light by use of CDC camera. We detected and compare the fluorescent light signal of INH resistant strains group (n= 16) and H37RV standard strains group (n= 10). The detection rate of codon 463 mutation accounted for 37% by comparing the above two groups. The coincidence rate of molecular beacon detection method and sequencing assay was 93 %. Conclusion The molecular beacon method is a technology with characteristics of high sensitiveness and specificity in detecting nucleic acid. The co-incidence rate is good between molecular beacon chip way and sequencing.
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