基因芯片技术诊断耐多药结核病的临床应用研究

发布时间：2018-03-10 07:45

本文选题：利福平　切入点：异烟肼　出处：《天津医药》2016年09期 　论文类型：期刊论文

【摘要】：目的探讨基因芯片法检测耐多药结核分枝杆菌临床分离株耐药相关基因rpo B、kat G和inh A突变的特点,评估其临床应用价值。方法选取2013—2014年石家庄地区耐多药结核分枝杆菌临床分离株76株,采用基因芯片法检测rpo B、kat G及inh A基因耐药突变位点及频率,以比例法药敏结果为金标准,评价基因芯片法检测菌株耐药的准确度、灵敏度和特异度,Kappa检验评价基因芯片法和比例法药敏结果的一致性。结果在76株石家庄地区耐多药结核分枝杆菌临床分离株中,69株检测到kat G/inh A基因发生单一或联合突变,其中kat G基因优势突变位点为315,占89.9%(62/69),5.8%(4/69)存在inh A-15(C→T)突变,4.3%(3/69)发生kat G 315位点和inh A启动子区域的联合突变。73株检测到rpo B基因发生突变,优势突变位点为531,占64.4%(47/73);其次是526,突变率为15.1%(11/73);12.3%(9/73)发生516位点突变;1.4%(1/73)发生513位点突变;1.4%(1/73)发生533位点突变;另有5.5%(4/73)发生多位点的联合突变。以比例法药敏结果为金标准,基因芯片法检测利福平耐药的敏感度为96.1%(73/76),特异度为100.0%(50/50),总准确度为97.6%(123/126);检测异烟肼耐药的敏感度为90.8%(69/76),特异度为100.0%(50/50),总准确度为94.4%(119/126);基因芯片法检测耐多药结核的敏感度为86.8%(66/76),特异度为100.0%(50/50),总准确度为92.1%(116/126)。结论本地区耐多药结核分枝杆菌耐药相关基因优势突变类型为kat G基因315位点和rpo B基因531位点,基因芯片法可快速、有效诊断耐多药结核病。
[Abstract]:Objective to investigate the characteristics of multidrug resistant Mycobacterium tuberculosis (MDR-M) clinical isolates using gene chip method to detect the mutations of drug-resistant related genes rpo Bnkat G and inh A. Methods 76 clinical isolates of Mycobacterium tuberculosis in Shijiazhuang area from 2013 to 2014 were selected to detect the mutation sites and frequencies of drug resistance in rpo Bu Kat G and inh A genes by gene chip assay. To evaluate the accuracy of gene chip method in the detection of drug resistance of strains, the results of proportional drug sensitivity were used as gold standard. The sensitivity and specificity of Kappa test were consistent with those of gene chip assay and proportional method. Results 69 of 76 clinical isolates of multidrug resistant Mycobacterium tuberculosis in Shijiazhuang were found to have a single or combined mutation of kat Gr / inh A gene. The dominant mutation site of kat G gene is 315, accounting for 89.9%. 鈫扵he mutation of rpo B gene was detected in the co-mutation of kat G315 locus and inh A promoter region. The dominant mutation locus was 531, accounting for 64.40.47 / 73; the second was 526, with a mutation rate of 15.11.11 / 73 / 12.30.The 516 locus mutation occurred 516 locus mutation 1.4 / 73) 533 locus mutation occurred, and 533 locus mutation occurred at 5.54r73). The results of proportional drug sensitivity were used as the gold standard. The sensitivity of gene chip assay for rifampicin resistance was 96. 1 / 73 / 76, the specificity was 10. 0 / 50 / 50, the total accuracy was 97. 6 / 123 / 126; the sensitivity for detecting isoniazid resistance was 90.8 / 69 / 76, the specificity was 10. 0 / 50 / 50 and the total accuracy was 94. 44 / 119 / 126; the sensitivity of gene chip method for multidrug resistant tuberculosis was 94. 4 / 19 / 126. The specificity was 100.050 / 50 and the total accuracy was 92.1a / 116 / 1260.Conclusion the dominant mutation types of drug-resistance related genes of Mycobacterium tuberculosis in this area are kat G gene 315 locus and rpo B gene 531 locus. Gene chip method is rapid and effective in the diagnosis of multi-drug resistant tuberculosis.
【作者单位】：石家庄市第五医院检验科;
【基金】：河北省卫生厅计划性课题(20150155)
【分类号】：R52;R440

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