摘要
目的利用焦磷酸测序技术,建立结核分枝杆菌菌株耐多药检测方法,并与Bactec MGIT 960药敏法进行比较。方法设计rpoB、katG基因PCR扩增引物和焦磷酸测序引物,建立结核分枝杆菌菌株焦磷酸测序检测方法。利用新确立的焦磷酸测序法检测202株临床分离的耐多药结核分枝杆菌的利福平、异烟肼耐药性。结果与BACTEC MGIT 960法检测药物耐药性结果比较,焦磷酸测序检测多耐药结核分枝杆菌临床分离株的敏感性与特异性为72.8%[95%可信区间（confidence in-terval,CI）：66.3%~78.4%]及90.0%（95%CI：80.1%~95.1%）。结论新确立的焦磷酸测序技术可快速检测结核分枝杆菌对利福平、异烟肼的耐药性,其可作为耐多药结核分枝杆菌药敏有效的检测工具。
Objective To develop the method of detecting multidrug-resistant Mycobacterium tuberculosis（MDR-TB） by pyrosequencing technology,and compare it with Bactec MGIT 960 susceptibility method.Methods The primers of the rpoB and katG genes were designed for PCR amplification and pyrosequencing and the pyrosequencing method was established for detection of Mycobacterium tuberculosis strains.The rifampicin and isoniazid resistances in 202 clinical isolates of MDR-TB were detected using the newly established pyrosequencing assay.Results Compared with BACTEC MGIT 960,the sensitivity and specificity of the pyrosequencing assay for detection of MDR-TB clinical isolates were 72.8% [95% confidence interval（confidence interval,CI）： 66.3-78.4%] and 90.0%（95% CI： 80.1-95.1%）,respectively.Conclusion The newly established pyrosequencing method can rapidly detect the resistances of Mycobacterium tuberculosis to rifampin and isoniazid,which can be used as a practical diagnostic tool for testing the resistance of MDR-TB.
出处
《实验与检验医学》
CAS
2012年第6期528-530,共3页
Experimental and Laboratory Medicine
基金
上海市科委项目(NO.10411955100)
江苏省卫生厅预防医学科研项目(Y2012095)
关键词
焦磷酸测序
检测
多耐药
结核
Pyrophosphate sequencing
Detection
Multidrug resistance
Tuberculosis
