DNA微阵列芯片技术检测培阳肺结核患者痰样本中结核分枝杆菌耐药性的价值  

Evaluation of the performance of DNA microarrays in detecting MTB drug resistance in sputum samples from patients with culture-positive pulmonary tuberculosis

作　　者：王悦[1] 孙颖 孙炳奇[1] WANG Yue;SUN Ying;SUN Bing-qi(Shenyang 10th People’s Hospital,Shenyang 110044,China)

机构地区：[1]沈阳市第十人民医院,沈阳110044

出　　处：《中国人兽共患病学报》2021年第2期109-114,共6页Chinese Journal of Zoonoses

基　　金：国家科技重大专项(No.2018ZX10103001-004-002)资助。

摘　　要：目的评价DNA微阵列芯片法(以下称芯片法)检测培养阳性患者痰样本的利福平和异烟肼相关耐药基因的效能。方法收集并检测389例疑似肺结核患者痰样,以BACTEC MGIT 960液体药敏(以下称MGIT 960药敏)为参考标准,评价芯片法检测样本利福平(RIF)、异烟肼(INH)耐药性和MDR-TB的灵敏度、特异度及一致性。应用线性探针耐药技术(GenoType MTBDRplus VER 2,0)对不一致的结果进行分析。结果将334例(334/389)MGIT 960培阳结果对应的痰处理液进行芯片法耐药检测,其中2例为非结核分枝杆菌,9例为阴性。与参考标准对比,芯片法对利福平耐药检测的灵敏度为92.4%,特异度为97.7%,Kappa值为0.90;异烟肼为82.6%,特异度为99.2%,Kappa值为0.86。在MDR-TB的病人中,芯片法与MGIT 960药敏法有较好的一致性;且在初治患者中Kappa值为0.95,灵敏度为95.5%,明显优于复治患者。结论线性探针耐药技术对22份不一致结果进行分析,认为检测限及耐药机制的差异、异质性耐药和多重感染等原因是导致基因型和表型或是两种分子法结果不一致的主要原因。应用芯片法检测培阳患者痰样本的利福平和异烟肼的相关耐药基因,与MGIT 960药敏法相比较具有较好的灵敏度、特异度和一致性,可快速检测出患者耐药情况和MDR-TB患者,为临床患者尤其是初治患者提供了快速有效的化疗参考方案。To evaluate the efficiency of a DNA microarray chip assay in detecting rifampicin and isoniazid drug resistance genes in sputum samples from patients with tuberculosis cultures,we collected and detected sputum samples from 389 patients with suspected pulmonary tuberculosis.The sensitivity,specificity and consistency of the chip method in detecting drug resistance to RIF and INH,and MDR-TB were evaluated,with the MGIT 960 test as the reference standard.If the results were inconsistent with those of the reference standard,the GenoType MTBDRplus technique from the WHO was used for verification.Drug resistance was detected with the chip method in 334 sputum samples with positive results detected by liquid culture,including two NTM positive cases and nine negative cases.Compared with those of the reference standard,the chip method’s sensitivity,specificity and Kappa value were 92.4%,97.7%and 0.90,respectively,in the rifampicin resistance test,and 82.6%,99.2%and 0.86,respectively,in the isoniazid resistance test.In patients with MDR-TB,the results were consistent with those of the liquid drug sensitivity test.In the new cases,the Kappa value was 0.95,and the sensitivity was 95.5%,values significantly better than those of the previously treated cases.In addition,22 inconsistent results were analyzed with GenoType MTBDRplus ver 2.0,it is considered that the detection limit and drug resistance mechanism,heterogeneous drug resistance and multiple infection are the main reasons for the inconsistent results of genotype and phenotype or two molecular methods.The chip method for the detection of rifampicin and isoniazid resistance genes in sputum samples of patients with positive cultures has good sensitivity,specificity and consistency,and can quickly detect resistance and MDR-TB.This method provides a rapid and effective chemotherapy reference scheme for clinical patients,particularly new cases.

关 键 词：结核分枝杆菌 DNA微阵列芯片法 MDR-TB 耐药 

分 类 号：R378.91[医药卫生—病原生物学]