基因芯片技术在雅安市结核分枝杆菌耐药检测中的应用

Application of Gene Chip Technology in the Detection of Mycobacterium Tuberculosis Resistance in Ya’an

目的评价基因芯片技术在雅安市结核分枝杆菌耐药检测工作中的应用价值。方法收集2016-06/2018-11雅安市结核病可疑患者痰培养分离株190株进行基因芯片菌种鉴定、耐药检测和比例法药敏试验。以比例法药物敏感试验为金标准,分析基因芯片技术检测结核分枝杆菌对利福平、异烟肼和耐多药诊断的敏感度、特异度、正确指数。采用McNemar检验比较两方法有无差异;采用Kappa检验比较两方法的一致性。两方法有差异的部分样本用基因测序的方法进行比对。结果基因芯片技术进行菌种鉴定,190株分离株中,有186株为结核分枝杆菌,4株为非结核分枝杆菌。以传统比例法药敏试验为金标准,186株结核分枝杆菌对利福平检测的敏感度和特异度为90.0%和96.6%,正确指数为0.866;对异烟肼检测的敏感度和特异度分别为89.5%和99.4%,正确指数为0.889;对耐多药的敏感度和特异度分别75.0%和99.4%,正确指数0.744。两种检测方法比较差异均无统计学意义(McNemar检验,P值分别为0.125、1.000和1.000)。两种检测方法具有较高的一致性(Kappa值分别为0.701、0.910、0.792)。基因芯片技术检测利福平耐药相关rpoB基因突变率最高的位点为531;异烟肼耐药相关katG基因主要突变位点为315。结论基因芯片技术与比例法药敏试验具有较好的一致性,灵敏度高、特异性好,是一种值得推广的耐药结核病诊断方法,在地市级结核病检测中具有重要的应用价值。

Objective To evaluate the value of the application of gene chip technology in the detection of Mycobacterium tuberculosis resistance in Ya’an. Methods From June 2016 to November2018,190 strains of suspected tuberculosis patients in Ya’an were collected for gene chip detection and proportional drug sensitivity test. The sensitivity,specificity and correct index of Mycobacterium tuberculosis for the diagnosis of rifampicin, isoniazid and multidrug resistance were analyzed by gene chip technique with the proportional method drug sensitivity test as the gold standard. The McNemar test was used to compare the differences between the two methods;the Kappa test was used to compare the consistency of the two methods. Some samples showing difference in different methods were compared by gene sequencing. Results Among the 190 strains, 186 strains were Mycobacterium tuberculosis and 4 strains were non-tuberculous mycobacteria. The traditional proportional method susceptibility test was taken as the gold standard, the sensitivity and the specificity of the 186 strains for rifampicin detection were 90.0% and 96.6%,and the correct index was and the correct index was 0.889. The sensitivity and the specificity for multidrug-resistant drugs were 75.0% and 99.4%, respectively, and the correct index was 0.744. There was no statistically significant difference between the two methods(McNemar test, P=0.125, 1.000, 1.000, respectively). The two detection methods had high consistency(Kappa=0.701, 0.910, 0.792, respectively). The site with highest mutation rate of rpoB gene related to rifampicin resistance was 531 by gene chip technology,and the main mutation site of katG gene related to isoniazid resistance was 315. Conclusion Gene chip technology and proportional susceptibility test have good consistency with high sensitivity and good specificity. It is a kind of drug-resistant tuberculosis diagnosis method worthy of promotion. It has important application value in the prefecture-level tuberculosis detection.