PCR-SSCP技术直接快速检测痰标本中结核分支杆菌rpoB,katG,ahpC基因突变的研究

Detection of rpoB, katG and ahpC genic mutation of mycobaterium tuberculosis in sputum specimens by PCR-SSCP

目的应用PCR -SSCP技术直接快速检测痰标本中结核分支杆菌rpoB ,katG及ahpC基因突变 ,评价此方法用于结核分支杆菌利福平 (RFP)及异烟肼 (INH)耐药性试验的意义。方法以结核分支杆菌H3 7Rv为对照 ,30例耐RFP(单耐或多耐 )、2 1例耐INH(单耐和多耐 )的结核分支杆菌临床分离株及 10例敏感菌株 ;39例菌阳肺结核患者及 30例非结核性肺部疾病患者痰标本 ,采用PCR SS CP技术检测痰标本和菌株中的结核杆菌rpoB ,katG ,ahpC基因突变 ,并与药敏试验对照。结果PCR -SSCP检测痰标本中结核分支杆菌rpoB ,katG及ahpC基因突变的阳性率分别为 79% (31/ 39)、46 % (18/39)及 5 % (2 / 39)。结论PCR -SSCP可望成为直接检测临床痰标本中结核分支杆菌耐利福平及耐异烟肼的快速方法。

Objective To evaluate the value of detection of rpoB, katG and ahpC genic mutation in Mycobacterium tuberculosis of sputums by PCR SSCP. Methods 30 cases clinical isolates of rifampin resistant and 21 cases clinical isolates of INH resistance Mycobacterium tuberculosis, 10 sensitive strains,39 sputum specimens of active pulmonary tuberculous patients and 30 sputem specimens from non tuberculous patients of pulmonary disease, with H37Rv reference as control group were tested by PCR SSCP. Results Of rpoB, katG and ahpC gene of sputum specimens the positive rate were 79%?46% and 5% respectively. Conclusion PCR SSCP may become a rapid method of direct detection of RFP resistat and INH resistance of Mycobacterium tuberculosis in clinical specimens.