煤工尘肺结核病耐药性的研究

Study on drug-resistance of Mycobacterium tuberculosis isolated from coal workeris with silico-tuberculosis

目的 了解煤工尘肺结核患者所染结核杆菌对异烟肼、利福平及链霉素耐药情况和基因突变情况 ,以便探寻更有效的药物治疗方法。方法 从 96份尘肺结核患者痰标本中分离出结核杆菌 ,测定异烟肼、利福平及链霉素的耐药情况 ,并用聚合酶链反应 -单链构象多态性分析法 (PCR SSCP)扩增出KatG、rpoB及rpsL片段 ,再将这些片段的 8%的非变性聚丙烯酰胺凝胶电泳图与标准株的电泳图进行对照分析。结果 常规药敏试验检测出各种耐药株共 6 7株 ,其中链霉素、利福平和异烟肼的耐药株构成比分别为 80 .5 %(5 4/ 6 7)、5 8.2 %(39/ 6 7)和 5 0 .7%(34 / 6 7) ;经PCR SSCP法分析 ,6 6株(98.5 %)rpsL泳动异常 ,47株 (70 .1%)出现rpoB泳动异常 ,42株 (6 2 .7%)KatG泳动异常。结论 大部分尘肺结核患者的结核杆菌耐药分离株有基因突变 ,其突变率高于常规药敏试验结果。

Objective To study the gene mutation and streptomycin,isoniazid or rifampicin resistance of Mycobacterium isolated from silico-tuberculosis patient's sputum so as to find a more effective therapy for this disease. Methods Mycobacteria tuberculosis were separated from 96 coal worker with silico-tuberculosis firstly.Then rpsL,KatG and rpoB fragments of genome were copied with PCR and compared their SSCP profiles with standard strains. Results 67 strains of streptomycin,isoniazid or rifampicin resistant Mycobacteria tuberculosis were found in routine drug resistence test,with the percentages of 80.5%(54/67),58.2%(39/67) and 50.7%(34/67) respectively.PCR-SSCP showed that out of 67 drug-resistant strains,66(98.5%) of rpsL,47(70.1%) of rpoB and 42(62.7%) of KatG appeared abnormal. Conclusion Most of the resistant strains appeared gene mutation.The mution rates were higher than the results from routine drug resistance test.