摘要
目的分析和探讨荧光PCR熔解曲线法对结核病的诊断和耐药检测的应用价值。方法收集2015年9月至2016年12月新疆维吾尔自治区胸科医院的976例疑似结核病患者的痰标本，运用探针荧光PCR熔解曲线法来检测结核分枝杆菌及其对利福平、异烟肼的耐药突变情况。以BACTECMGIT960液体培养法及液体药敏试验检测结果为标准，评价探针荧光PCR熔解曲线法鉴定结核分枝杆菌及其耐药突变检测的敏感度、特异度、一致率、Kappa值等。结果以MGIT960液体培养结果为标准，荧光PCR熔解曲线法鉴定结核DNA的敏感度为85．44％（135／158），特异度为94．01％（769／818），Kappa值为0．75，诊断符合率为92．62％（904／976）。以MGIT960液体药敏试验结果为标准，荧光PCR熔解曲线法对异烟肼耐药突变检测的敏感度为83．33％（20／24），特异度为94．59％（105／111），Kappa值为0．76，诊断符合率为92．59％（125／135）；荧光PCR熔解曲线法对利福平耐药突变检测的敏感度为95．83％（23／24），特异度为95．50％（106／111），Kappa值为0．86，诊断符合率为95．56％（129／135）。结论荧光PCR熔解曲线法检测速度快，对利福平和异烟肼耐药突变检测结果具有良好的敏感度和特异度，可用于临床上对结核分枝杆菌利福平和异烟肼耐药情况的快速筛查。
Objective To evaluate the value of fluorescence PCR melting curve analysis in Mycobacterium tuberculosis （MTB） identification and drug-resistant TB detection. Methods The sputum specimens from 976 sus- pected TB patients were collected during September 2015 to December 2016 in Chest Hospital of Xinjiang Uyghur Autonomous Region. MTB and its drug resistance to rifampiein and isoniazid were detected by fluorescence PCR melting curve analysis. The results of MGIT 960 liquid culture method and liquid drug resistance test were stipulated as criterion to evaluate the sensitivity, specificity, agreement rate and Kappa value of melting curve analysis in MTB identification and drug-resistant TB detection. Results The sensitivity, specificity, Kappa value and diagnostic coincidence rate of the melting curve analysis for MTB DNA identification were 85.44% （135/158）, 94.01% （769/818）, 0.75 and 92. 620/00 （904/976）, respectively, taking the result of MGIT 960 liquid culture as gold standard. Taking MGIT 960 liquid drug resistance test as gold standard, the sensitivity, specificity, Kappa value and diagnostic coincidence rate of the melting curve analysis for isoniazid-resistant mutation detection were 83.33% （20/24）, 94.59% （105/111）, 0.76 and 92.59% （125/135）, respectively. Meanwhile, the sensitivity, specificity, Kappa value and diagnostic coincidence rate of the melting curve analysis for rifampin-resistant mutation detection were 95.83% （23/24）, 95.50% （106/111）, 0. 86 and 95.56% （129/135）, respectively. Conclusion The probe based melting curve analysis can detect rifampicin-resistant and isoniazid-resistant mutation rapidly and accurately, which can be used for the rapid detection of the multi-drug resistant mutations of MTB.
作者
刘艳
古丽比克·木拉提
玛力亚木·阿布力提甫
易星
李君莲
师永欢
刘天剑
LIU Yan;Gulibike·Mulati;Maliyamu·Abulitifu;YI Xing;LIJun-lian;SHI Yong-huan;LIU Tian-jian(Department of Clinical Laboratory,Chest Hospital of Xinjiang Uyghur Autonomous Region,Urumq 830049,China)
出处
《中国防痨杂志》
CAS
2018年第9期959-963,共5页
Chinese Journal of Antituberculosis
