DNA微陈列芯片法快速检测结核分枝杆菌rpoB、katG和inhA基因突变的临床应用

Clinical application of rapid detection of rpoB,katG and inhA gene mutations in Mycobacterium tuberculosis by DNA micro-display chip method

目的:探讨DNA微陈列芯片法快速检测结核分枝杆菌rpoB、katG和inhA基因突变的临床应用效果。方法:通过数字表法选择2014年5月—2016年5月初步诊断为结核病患者70例的痰样,使用DNA微陈列芯片法快速检测,并同时通过传统药敏试验和DNA测序法进行再次检查以作为对照。结果:70株结核分枝杆菌中药物敏感株芯片杂交共计20株,这一结果完全符合标准株,符合率100%(20/20),30株耐利福平(RIF)临床分离株均检测到rpoB基因突变,基因突变检出率为100%(30/30),50株异烟肼耐药株中有41株检测katG基因突变,检出率82.00%,9株检测到inhA基因突变,检出率18.00%(9/50),这一检测结果和测序结果完全符合。结论:DNA微陈列芯片法快速检测结核分枝杆菌rpoB、katG和inhA基因突变效果良好,虽然暂且不能代替传统的药敏试验方法,但其检测的灵敏度高,特异性和重复性好。

Objective： To investigate the clinical effect of rapid detection of rpoB,katG and inhA gene mutations of Mycobacterium tuberculosis by DNA micro-display method. Methods： Through the digital meter method selection from May 2014 to May 2016 in hospital during the initial diagnosis of tuberculosis patients collected 70 cases of sputum samples,using DNA micro display chip method of fast detection,and at the same time through the traditional drug susceptibility testing and DNA sequencing method to check again as to explore the contrast analysis. Results： All 70 strains of Mycobacterium tuberculosis in drug sensitive strains hybridization totaling 20 strains,the results accord with the standard strain,the coincidence rate was100%（ 20/20）,RIF（ Li Fuping） resistance in 30 strains of clinical isolates were detected rpoB gene mutation,gene mutation rate was 100%（ 30/30）,50 strains of isoniazid resistant strains there are 41 strains of katG gene mutation detection,the detection rate of 82. 00%,9 strains were detected inhA gene mutation detection rate of 18. 00%（ 9/50）,and the test results and the sequencing result is entirely consistent. Conclusion： The method for rapid detection of mycobacterium tuberculosis DNA micro display chip rpoB,katG,and inhA gene mutation effect is good,though unable to replace the traditional drug sensitive test method,but its high detection sensitivity,specificity and good repeatability.