Email Alert | RSS    帮助

中国防痨杂志 ›› 2019, Vol. 41 ›› Issue (1): 74-79.doi: 10.3969/j.issn.1000-6621.2019.01.016

• 论著 • 上一篇    下一篇

荧光PCR探针熔解曲线法检测结核分枝杆菌复合群对利福平和异烟肼耐药性的价值

吴慧娜1,孙付胜2(),刘清文1,戎中录1,张纪东1,高阳1,李永福1,常斐1   

  1. 1. 274000 山东省菏泽市传染病医院检验科,感染科
    2. 山东省菏泽市疾病预防控制中心结核病防治科
  • 收稿日期:2018-11-05 出版日期:2019-01-10 发布日期:2019-01-09
  • 基金资助:
    山东省医药卫生科技发展计划(2016WS0090)

The value of fluorescent PCR probe melting curve analysis in detection of Mycobacterium tuberculosis complex for rifa-mpicin and isoniazid resistance

Hui-na WU1,Fu-sheng SUN2(),Qing-wen LIU1,Zhong-lu RONG1,Ji-dong ZHANG1,Yang GAO1,Yong-fu LI1,Fei CHANG1   

  1. 1. Department of Clinical Laboratory, Heze City Infectious Diseases Hospital, Shandong Province, Heze 274000, Chin
  • Received:2018-11-05 Online:2019-01-10 Published:2019-01-09

摘要:

目的 评价荧光PCR探针熔解曲线法(简称“探针熔解曲线法”)检测结核分枝杆菌复合群对利福平(RFP)和异烟肼(INH)耐药性的价值。方法 从2015年1月至2018年7月山东省菏泽市传染病医院收治的883例肺结核和耐多药结核病(MDR-TB)患者中,选择萋-尼染色结果为阳性的结核病患者的痰标本共215份(例)进行分离培养,对鉴定为结核分枝杆菌复合群(MTBC)的200株(例)菌株同时使用比例法药物敏感性试验(DST)和探针熔解曲线法检测对RFP和INH的耐药性。以DST检测结果为金标准,分析探针熔解曲线法检测RFP和INH的敏感度、特异度、符合率和一致性(Kappa检验)。结果 200株(例)MTBC分离株中,以DST检测结果为标准,探针熔解曲线法检测RFP耐药性的敏感度、特异度和符合率分别为96.4%(106/110;95%CI:90.7%~98.9%)、80.0%(72/90;95%CI:70.5%~87.1%)和89.0%(178/200);对INH耐药性检测的敏感度、特异度和符合率分别为85.4%(123/144;95%CI:78.6%~90.7%)、96.4%(54/56;95%CI:87.7%~99.6%)和88.5%(177/200)。探针熔解曲线法与DST检测MTBC对RFP耐药性的Kappa值为0.78,对INH耐药性的Kappa值为0.74。结论 探针熔解曲线法检测MTBC对RFP和INH的耐药性均有较高的敏感度和特异度,且探针熔解曲线法与DST检测MTBC对RFP和INH耐药性的一致性较高,有助于对耐药结核病患者的及早发现和治疗。

关键词: 分枝杆菌, 结核, 聚合酶链反应, 分子探针技术, 利福平, 异烟肼, 微生物敏感性试验, 数据说明, 统计

Abstract:

Objective To evaluate the value of fluorescent PCR probe melting curve analysis (MeltProTB) in detecting the resistance of Mycobacterium tuberculosis complex to rifampicin (RFP) and isoniazid (INH).Methods Eight hundred and eighty-three pulmonary tuberculosis patients with multidrug-resistant tuberculosis (MDR-TB) admitted in Heze City Infectious Diseases Hospital in Shandong Province were enrolled during January 2015 to July 2018. Two hundred and fifteen patients with sputum smear positive were selected for further culture. Two hundred strains identified as Mycobacterium tuberculosis complex (MTBC) were tested for drug resistance to RFP and INH by proportional drug susceptibility testing (DST) and MeltPro TB method. The sensitivity, specificity, coincidence rate and consistency (Kappa test) of resistance to RFP and INH detected by MeltPro TB were analyzed using DST results as gold standard.Results Of the 200 MTBC isolates, the sensitivity, specificity and coincidence rate of RFP resistance were 96.4% (106/110; 95%CI: 90.7%-98.9%), 80.0% (72/90; 95%CI: 70.5%-87.1%) and 89.0%(178/200) by MeltPro TB based on DST as the standard. The sensitivity, specificity and coincidence rate of RFP resistance were 85.4% (123/144; 95%CI: 78.6%-90.7%), 96.4% (54/56; 95%CI: 87.7%-99.6%) and 88.5% (177/200) by MeltPro TB based on DST as the standard. The Kappa value of MeltPro TB and DST for RFP resistance in MTBC was 0.78, and was 0.74 for INH resistance in MTBC.Conclusion MeltPro TB has high sensitivity and specificity in detecting drug resistance of MTBC to RFP and INH, and it has good consistent with DST in detecting MTBC resistance to RFP and INH, which is helpful for early detection and treatment in patients with drug-resistant tuberculosis.

Key words: Mycobacterium tuberculosis, Polymerase chain reaction, Molecular probe techniques, Rifampin, Isoniazid, Microbial sensitivity tests, Data interpretation, statistical