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目的：研究结核分枝杆菌（Ｍｙｃｏｂａｃｔｅｒｉｕｍｔｕｂｅｒｃｕｌｏｓｉｓ，ＭＴ）耐药的分子机制，建立快速检测耐药基因型的分子药敏试验方法。方法：通过聚合酶链反应（Ｐｏｌｙｍｅｒａｓｅｃｈａｉｎｒｅａｃｔｉｏｎ，ＰＣＲ）、ＰＣＲ－单链构象多态性（Ｓｉｎ－ｇｌｅ－ｓｔｒａｎｄｅｄｃｏｎｆｏｒｍａｔｉｏｎｐｏｌｙｍｏｒｐｈｉｓｍ，ＳＳＣＰ）、ＰＣＲ－限制性片段长度多态性（Ｒｅｓｔｒｉｃｔｉｏｎｆｒａｇｍｅｎｔｌｅｎｇｔｈｐｏｌｙ－ｍｏｒｐｈｉｓｍ，ＲＦＬＰ）分析ＭＴ临床分离株的ｒｐｏＢ、ｒｐｓＬ、ＫａｔＧ基因和ｉｎｈＡ调节序列。结果：ＰＣＲ分析耐药基因的敏感性为１～１０ｐｇＤＮＡ；除ｒｐｏＢ引物ＰＣＲ扩增为属特异性外，余耐药基因引物ＰＣＲ扩增都具有较高的种特异性。与传统药敏试验方法相比，ＰＣＲ和ＰＣＲ－ＳＳＣＰ检测６２株ＭＴ分离株利福平（Ｒｉｆａｍｐｉｎ，ＲＦＰ）、链霉素（Ｓｔｒｅｐｔｏ－ｍｙｃｉｎ，ＳＭ）和异烟肼（Ｉｓｏｎｉａｚｉｄ，ＩＮＨ）耐药基因型的灵敏度和特异度分别为９３．８％和９６．２％（ｒｐｏＢ）、８１．６％和９１．７％（ｒｐｓＬ）、７６．０％和７５．７％（ｋａｔＧ）、１６％和１００％（ｉｎｈＡ）。?
Objective: To study the molecular mechanism of Mycobacterium tuberculosis (MT) resistance and to establish a molecular susceptibility test for rapid detection of drug resistance genotypes. Methods: MT clinical data were analyzed by polymerase chain reaction (PCR), PCR-SSCP and PCR-restriction fragment length polymorphism (RFLP) RpoB, rpsL, KatG genes and inhA regulatory sequences of isolates. Results: The sensitivity of PCR analysis of resistance genes was 1 ~ 10 pg DNA. Except for the PCR amplification of rpoB primers was genus specific, the PCR-amplified PCR primers were highly specific. Compared with traditional drug susceptibility testing methods, 62 MT isolates of Rifampin (RFP), Strepto-mycin (SM) and Isoniazid (INH) were detected by PCR and PCR- The sensitivity and specificity of the genotypes were 93.8% and 96.2% (rpoB), 81.6% and 91.7% (rpsL), 76.0% and 75.7% (katG), 16% And 100% (inhA). ?