PCR-反向点杂交法检测复治涂阳患者结核分枝杆菌耐药性的价值

目的 评估PCR-反向点杂交法检测复治涂阳患者痰标本中MTB耐药性的价值。方法 收集2015年6月至2019年1月上海市肺科医院诊治的400例复治涂阳肺结核患者的痰标本,每份标本量均不少于2ml;每例患者采用同一标本进行PCR-反向点杂交法MTB耐药基因检测、GeneXpert MTB/RIF检测和BACTEC MGIT 960液体培养、菌种鉴定、微孔板法药物敏感性试验(简称“药敏试验”),最终纳入357例(株)为研究对象。以微孔板法药敏试验结果为参考标准,评价PCR-反向点杂交法的检测效能。结果 以微孔板法药敏试验结果为参考标准,PCR-反向点杂交法检测MTB对利福平耐药性的敏感度、特异度、阳性预测值、阴性预测值、符合率和Kappa值分别为97.5%(115/118)、97.1%(232/239)、94.3%(115/122)、98.7%(232/235)、97.2%(347/357)和0.937,对异烟肼耐药性的敏感度、特异度、阳性预测值、阴性预测值、符合率和Kappa值分别为82.5%(113/137)、99.1%(218/220)、98.3%(113/115)、90.1%(218/242)、92.7%(331/357)和0.841,对链霉素耐药性的敏感度、特异度、阳性预测值、阴性预测值、符合率和Kappa值分别为86.5%(115/133)、99.1%(222/224)、98.3%(115/117)、92.5%(222/240)、94.4%(337/357)和0.877,对乙胺丁醇耐药性的敏感度、特异度、阳性预测值、阴性预测值、符合率和Kappa值分别为60.7%(37/61)、98.6%(292/296)、90.2%(37/41)、92.4%(292/316)、92.2%(329/357)和0.682。结论 PCR-反向点杂交法检测复治涂阳患者痰标本中MTB耐药性有较好的效能。

The value of PCR-reverse dot blot hybridization in detecting the drug resistance of Mycobacterium tuberculosis in sputum specimens of retreatment smear-positive pulmonary tuberculosis patients

Objective To investigate the value of PCR-reverse dot blot (RDB) hybridization in detecting the drug resistance of Mycobacterium tuberculosis(MTB) in sputum specimens of retreatment smear-positive pulmonary tuberculosis patients. Methods Sputum specimens from 400 retreatment smear-positive pulmonary tuberculosis patients diagnosed and treated in Shanghai Pulmonary Hospital from June 2015 to January 2019 were collected. The amount of each specimen was more than 2 ml. The same sputum specimen of each patient was tested for drug resistance detection using PCR-RDB, Gene Xpert MTB/RIF and Micropore-plate method (MicroDST). Three hundred and fifty-seven cases (strains) were finally included. The results of MicroDST were used as reference standard to evaluate the performance of PCR-RDB. Results Based on reference standard, the sensitivity, specificity, positive predictive value, negative predictive value, coincidence rate and Kappa value of PCR-RDB in detecting rifampicin resistance of MTB were 97.5% (115/118), 97.1% (232/239), 94.3% (115/122), 98.7% (232/235), 97.2% (347/357) and 0.937; the sensitivity, specificity, positive predictive value, negative predictive value, coincidence rate and Kappa value of PCR-RDB in detecting isoniazid resistance of MTB were 82.5% (113/137), 99.1% (218/220), 98.3% (113/115), 90.1% (218/242), 92.7% (331/357) and 0.841; the sensitivity, specificity, positive predictive value, negative predictive value, coincidence rate and Kappa value of PCR-RDB in detecting streptomycin resistance of MTB were 86.5% (115/133), 99.1% (222/224), 98.3% (115/117), 92.5% (222/240), 94.4% (337/357) and 0.877; the sensitivity, specificity, positive predictive value, negative predictive value, coincidence rate and Kappa value of PCR-RDB in detecting ethambutol resistance of MTB were 60.7% (37/61), 98.6% (292/296), 90.2% (37/41), 92.4% (292/316), 92.2% (329/357)and 0.682. Conclusion PCR-reverse dot blot hybridization shows a good capability in detecting the drug resistance of MTB in sputum specimen of retreatment smear-positive pulmonary tuberculosis patients.