结核分枝杆菌表型药敏与荧光PCR探针溶解曲线法检测一致性分析

Consistency analysis of phenotypic drug sensitivity detection of mycobacterium tuberculosis detection with fluorescence PCR probe dissolution curve method

目的比较结核分枝杆菌表型药敏与快速分子检测结果的一致性,探讨适合喀什地区耐药结核病筛查方法和检测技术,为耐药结核病的早期发现和临床诊断提供参考依据。方法从2022年1—12月喀什地区肺科医院确诊的结核病患者200份痰涂片阳性标本分离菌株,同时开展结核菌罗氏固体培养法(简称表型药敏)与荧光PCR探针溶解曲线法(简称分子药敏)试验,比较两种药敏试验结果的一致性。结果200份标本中,共检出结核分枝杆菌183株,对利福平药物敏感试验两种检测方法结果一致176株(敏感170株、耐药6株)、一致率96.17%;对异烟肼药物敏感试验两种检测方法结果一致174株(敏感167株、耐药7株)、一致率95.08%;分子药敏与表型药敏检测结核分枝杆菌耐药性的Kappa值对利福平为0.54、对异烟肼为0.49。利福平耐药菌株中,利福平ropB基因突变位点数10株,表型药敏利福平耐药突变位点数8株;异烟肼耐药菌株中,异烟肼katG、inhA基因突变位点数8株,表型药敏异烟肼耐药突变位点数7株。结论结核分枝杆菌表型药敏与快速分子检测药敏结果一致性较好,分子药敏技术可作为初筛诊断工具,有助于早期、快速筛查耐药结核病;建议喀什地区有条件的县(市)级结核病定点医院使用两种检测方法平行检测,以提高精准度,及早发现传染源,有效防止结核病疫情的蔓延。

Objective To compare the consistency of phenotypic drug sensitivity and rapid molecular detection results of mycobacterium tuberculosis,and explore suitable screening methods and detection techniques for drug-resistant tuberculosis in Kashi prefecture,so as to provide reference for early detection and clinical diagnosis of drug-resistant tuberculosis.Methods The strains were isolated from 200 sputum smear positive samples of tuberculosis patients diagnosed in pulmonary diseases hospital from January to December 2022.Meanwhile,the solid culture method(phenotypic drug sensitivity for short)and fluorescent PCR probe dissolution curve method(molecular drug sensitivity for short)were carried out to compare the consistency of the results of both drug sensitivity tests.Results A total of 183 strains of mycobacterium tuberculosis were detected in 200 samples.The results of both detection methods for rifampicin drug sensitivity test were consistent in 176 strains,of which,170 strains were sensitive and 6 strains were resistant,with the consistency rate of 96.17%.The results of both detection methods for isoniazid drug sensitivity test were consistent in 174 strains(167 strains were sensitive and 7 strains were resistant),with the consistency rate of 95.08%.The Kappa values for molecular and phenotypic drug sensitivity testing of mycobacterium tuberculosis resistance were 0.54 for rifampicin and 0.49 for isoniazid.Among the rifampicin-resistant strains,there were 10 rifampicin ropB gene mutations and 8 phenotypic drug sensitive rifampicin resistant mutations.Among the isoniazid resistant strains,there were 8 mutations in the katG and inhA genes of isoniazid,and 7 mutations in phenotypic drug sensitive isoniazid resistant strains.Conclusions The consistence of the results of phenotypic drug sensitivity detection of mycobacterium tuberculosis is good with the results of rapid molecular detection.Molecular drug sensitivity technology can be used as a preliminary screening diagnostic tool,which is helpful for early and rapid screening of drug-resistant tuberculosis.It is suggested that the pulmonary diseases hospitals in the qualified counties(cities)of Kashi prefecture could use two detection methods to improve the accuracy,and find the source of infection source as soon as possible,so as to effectively prevent the spread of tuberculosis epidemic.