目的研究荧光PCr熔解曲线法检测结核分枝杆菌耐异烟肼突变方法,评价检测耐多药结核病的能力。方法对结核病人临床分离结核分枝杆菌使用荧光PCr熔解曲线法检测,与金标准传统药物敏感性试验比较,获得该方法的灵敏度、特异度和一致率。结果检测408例结核病人临床分离培养样本,药敏试验检出278例异烟肼敏感标本,130例异烟肼耐药标本;荧光PCr探针熔解曲线法检出异烟肼敏感和耐药标本281例、127例,灵敏度为87.69%,特异性为95.32%,一致率为92.89%。结论荧光PCr探针熔解曲线法检测速度快速、特异性强、灵敏度较高,可用于结核分枝杆菌耐异烟肼突变的快速检测,适于耐多药结核病的快速筛查。OBJECTIVE The study explored the features of fluorescent PCR / melt-curves analysis in the detection of isoniazid-resistant mutations in Mycobacterium tuberculosis, so to evaluate the capability of the method in the identification of multidrug-resistant tuberculosis.METHODS Mycobacterium tuberculosis isolated from tuberculosis patients were subjected to fluorescent PCR / melt-curves analysis and gold standard conventional drug susceptibility test.The results were compared to obtain the sensitivity, specificity, and consistency of the new method.RESULTS Among the 408 culture samples isolated from tuberculosis patients, drug susceptibility test indicated 278 isoniazid-sensitive and 130 isoniazid-resistant specimens, and fluorescent PCR / melt-curves analysis revealed 281 and 127 specimens, respectively.Consequently, sensitivity, specificity,and consistency of the fluorescent PCR / melt-curves analysis were 87.69%, 95.32%, and 96.32%, respectively.CONCLUSION Fluorescent PCR / melt-curves analysis is time-efficient and high in specificity and sensitivity, and may be used in the rapid detection of isoniazid-resistant mutations in Mycobacterium tuberculosis and rapid screening of multidrug-resistant tuberculosis.“艾滋病和病毒性肝炎等重大传染病防治”科技重大专项资助项目(2008ZX10003-004
