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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 understand the resistance of Mycobacterium tuberculosis to isoniazid, rifampicin and streptomycin and the mutation of gene in patients with pneumoconiosis tuberculosis in order to explore more effective drug treatment. Methods Mycobacterium tuberculosis was isolated from sputum samples of 96 patients with pneumoconiosis. The drug resistance of isoniazid, rifampicin and streptomycin was determined. PCR-SSCP (polymerase chain reaction-single strand conformation polymorphism) The KatG, rpoB and rpsL fragments were amplified and compared with 8% non-denaturing polyacrylamide gel electrophoresis of these fragments and the standard strains electrophoresis. Results A total of 67 resistant strains were detected by routine susceptibility test. The resistant strains of streptomycin, rifampicin and isoniazid were 80.5% (5/67), 5 8.2 (39/67) and 50.7% (34/67) respectively. PCR-SSCP analysis showed that 66 (98.5%) rpsL migrated abnormally and 47 (70.1%) showed rpoB migration Abnormalities, 42 (62.7%) KatG motility abnormalities. Conclusion Most of the patients with TB pneumoconiosis have mutations in M. tuberculosis isolates, and their mutation rate is higher than that of conventional susceptibility test.