摘要
目的高分辨率熔解曲线(High resolution melting,HRM)技术检测结核菌耐药的结果评价。方法收集自2011年1月至2012年12月期间在深圳市第三人民医院住院进行抗痨治疗的结核病患者的痰标本134例,应用全自动快速分枝杆菌培养鉴定药敏系统BACTC-MGIT 960进行快速检测,获得所需要的结核菌培养药敏结果。针对利福平耐药相关rpo B基因的利福平耐药决定区区域和异烟肼耐药相关基因kat G315位点和inh A-15位点分别进行HRM扩增分析,从而获得利福平耐药决定区、kat G315、inh A-15的基因突变信息,进而判断各菌株是否存在利福平和异烟肼的耐药。结果 134株结核菌临床分离株经结核菌药敏实验判定,有28株对利福平耐药,27株对异烟肼耐药,23株同时对利福平和异烟肼耐药。以结核菌药敏结果为金标准,HRM检测利福平耐药敏感性为96.4%(27/28),特异性为97.2%(103/106),阳性预测值为90.0%(27/30),阴性预测值为99.0%(103/104);异烟肼耐药敏感性为88.9%(23/27),特异性为97.6%(101/107),阳性预测值为79.3%(23/29),阴性预测值为96.2%(101/105)。结论 HRM快速检测结核菌耐药,具有较高的特异性、灵敏度、阳性预测值和阴性预测值,能够满足临床需求。
Objective To assess result of detection of Mycobacterium tuberculosis resistance by high resolution melting analysis. Methods Total 134 sputum from tuberculosis patients in the Third People＇ s Hospital of Shenzhen from 2011 to 2012 were collected. Drug susceptibility testing was done by BACTC--MGIT 960 system, and high resolution melting analysis was developed to scan for mutations in the rpoB, katG, inhA genes for the detection of rifampin and isoniazid resistance of Mycobaeterium tuberculosis. High resolution melting analysis was performed for the rifampin resistance-determining region of the rpoB as well as isoniazid resistance-related genes katG315 and inhA-15. Correspondenfly, genetic mutation information of these loci was obtained to determine whether drug resistance to rifampin or isoniazid was presented in each of the isolates. Results A total of 134 clinical MTB strains were tested by BACTC--MGIT 960 system, and there were 28 RIF-resistant,27 INH resistant, 23 MDR-TB. Compared against results of drug susceptibility testing, for detecting rifampin resistance, the sensitivity and specificity of HRM were 96.4% （27/28） and 97.2% （103/106）, the positive predictive value and negative predictive value of HRM were 90%（27/30） and 99%（103/104）, for detecting isoniazid resistance, the sensitivity and specificity of HRM were 88.9% （23/27） and 97.6% （101/107）,the positive predictive value and negative predictive value of HRM were 79.3% （23/29） and 96.2% （101/105）. Conclusion Detection of multidrug-resistant Myeobacterium tuberculosis by high resolution melting analysis can meet for clinical demand.
出处
《中国热带医学》
CAS
2014年第12期1435-1439,共5页
China Tropical Medicine
基金
广东省新发传染病诊治重点实验室项目资助(No.2011A060901009)
关键词
高分辨率熔解曲线技术
结核
耐药
High resolution melting
Tuberculosis
Drug-resistance
