DNA微阵列芯片法检测结核分枝杆菌耐异烟肼基因的研究

目的采用DNA微阵列芯片法检测结核分枝杆菌INH耐药基因,为临床诊治提供实验依据。方法应用DNA微阵列芯片法和改良罗氏培养加比例法药敏试验对疑似结核病患者的150份痰液标本进行检测,并对检测的结果进行比较分析。结果 150份痰液标本的涂阳率为60.0%(90/150),DNA微阵列芯片法检测到结核分枝杆菌阳性率为62.7%(94/150),改良罗氏培养法阳性率为60.0%(90/150),比例法药敏试验异烟肼的耐药率为43.9%(36/82),异烟肼的kat G/inh A基因总突变率为35.1%(33/94);以改良罗氏培养加比例法药敏试验为标准,DNA微阵列芯片法检测结核分枝杆菌阳性、异烟肼耐药性和敏感性的符合率分别为97.7%、86.1%和100.0%。结论 DNA微阵列法可快速准确地检测大部分疑似结核病临床标本的kat G和inh A基因突变,可用于临床耐药性的检测从而指导临床用药,也可用于临床诊断中的推广。

The study of DNA microarray chip method to test the resistance isoniazid gene of Mycobacterium tuberculosis

Objective To test the resistance isoniazid gene of Mycobacterium tuberculosis using the method of DNA microarray chip.Methods A total of 150 sputum specimens of suspected tuberculosis (TB) patients were tested by the method of DNA microarray chip and the modified Roche medium proportion method,and the results were analyzed and compared.Results The rate of sputum smear positive was 60.0％(90/150) among the 150 specimens.The positive rate of Mycobacterium tuberculosis was 62.7％(94/150) by the DNA microarray chip method,and the positive rate was 60.0％(90/150)by the modified Roche medium proportion method.The drug resistance rate of isoniazid was 43.9％(36/82) by the drug sensitive test of the proportion method.The katG/inhA gene mutation rate of isoniazid was 35.1％(33/94).With the standard of modified Roche medium proportion method,the coincidence rates of the Mycobacterium tuberculosis positive rate,isoniazid drug resistance rate,and isoniazid drug sensitive rate tested by the DNA microarray chip were 97.7％,86.1％ and 100.0％ respectively.Conclusion The DNA microarray method can quickly and reliably detect the gene mutation of katG and inhA in most clinical samples of suspected TB patients.This method can be used to detect clinical drug resistance and guide clinical medication and diagnosis.