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  

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

作　　者：刘轾彬[1] 吴敏 吴小翠[2] 韩敏[2] 肖和平[1] 张青[1] LIU Zhi-bin;WU Min;WU Xiao-cui;HAN Min;XIAO He-ping;ZHANG Qing(Department of Tuberculosis,Shanghai Pulmonary Hospital Affiliated to Tongji University School of Medicine,Shanghai 200433,China)

机构地区：[1]同济大学附属上海市肺科医院结核科,200433 [2]同济大学附属上海市肺科医院检验科,200433

出　　处：《中国防痨杂志》2021年第1期47-51,共5页Chinese Journal of Antituberculosis

基　　金：“十三五”国家科技重大专项(2018ZX10722302-002);上海市感染性疾病(结核病)临床医学研究中心项目(19MC1910800)。

摘　　要：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 MGIT960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。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 Kappavalue 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 Kappavalue 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.

关 键 词：分枝杆菌 结核 结核 抗多种药物性 微生物敏感性试验 聚合酶链反应 双杂交系统技术 评价研究 数据说明 统计 

分 类 号：R52[医药卫生—内科学] R446.5[医药卫生—临床医学]