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目的通过诱导试验观察结核菌产生耐利福平药物的全过程,初步分析结核菌耐该药的机理。方法采用诱导试验、聚合酶链反应—单链构象多态性(PCR-SSCP)和序列分析,对经不同药物浓度传代培养后的结核菌强毒株(H_(37)Rv)和36株临床耐药分离株进行分析。结果在诱导到第7代时,H_(37)Rv的PCR-SSCP电泳出现异常,经测序证实在513位点发生基因突变。36株临床耐药分离株中有23株发生突变,为63.9%(23/36);其中有5株与诱导株基因突变位点相同。结论用药物不间断的刺激结核菌,是产生结核菌耐利福平药物的重要原因。
Objective To observe the whole course of rifampicin-producing drug produced by Mycobacterium tuberculosis by induction test and to analyze the mechanism of tuberculosis-resistant drug. Methods The virulent strains of Mycobacterium tuberculosis (H_ (37) Rv) and the clinical isolates of 36 clinical isolates after subculture with different drug concentrations were detected by induction test, polymerase chain reaction and single strand conformation polymorphism (PCR-SSCP) Resistant isolates were analyzed. Results PCR-SSCP electrophoresis of H_ (37) Rv was abnormal at the 7th generation, and the gene mutation was confirmed at the 513 site by sequencing. Twenty-three out of 36 clinical isolates were mutated, accounting for 63.9% (23/36). Five of them were the same as the induced mutants. Conclusion The uninterrupted stimulation of drug with Mycobacterium tuberculosis is an important reason for the production of rifampicine in tuberculosis.