【摘要】 目的 探讨利用分子生物学方法直接快速检测临床标本中淋球菌gyrA基因突变的可行性。 方法 设计针对淋球菌gyrA基因的特异扩增引物，运用聚合酶链反应-单链构象多态性技术(PCR-SSCP)对95份泌尿生殖道标本直接进行基因及突变检测，并与测序及药敏试验结果进行对比分析。 结果 32份分离培养阴性的标本无特异性扩增产物，而63份分离培养阳性标本以及对应的分离菌株均能扩增出168bp片断。以淋球菌标准株ATCC19424为对照，分离菌株对环丙沙星敏感的5份临床标本与标准株SSCP带型相同;分离菌株对环丙沙星中介或耐药的58份临床标本与标准株SSCP带型均不同，共出现5种突变带型。5份SSCP分析显示，无突变的标本gyrA基因序列未发生改变;16份SSCP分析显示，有突变的标本gyrA基因序列均有改变;具有相同突变的标本SSCP带型完全相同，具有不同突变类型的标本SSCP带型不同。 结论 PCR-SSCP银染技术可简便、快速、准确地检测临床标本中淋球菌对氟喹诺酮类药物的敏感性。

    【关键词】 喹诺酮类;DNA突变分析

    【Abstract】 Objective To investigate the feasibility of directly detecting the gyrA gene mutation of Neisseria gonorrhoea by molecular biotechnology in clinical specimens.Methods The gyrA gene was amplified by PCR in95clinical specimens.The PCR products were detected by single strand conforma-tion polymorphism(SSCP)and directly sequenced.The results were compared with those of drug sus-ceptibility test and sequencing analysis.Results The168bp fragments could be amplified from63posi-tive specimens and corresponding isolated strains.There were no specific amplified products in32neag-tive specimens.SSCP patterns in5specimens whose corresponding isolates were susceptible to ciprofloxacin were the same with wild type strains.SSCP patterns in58specimens whose corresponding isolates were intermediate or resistant to ciprofloxacin were different from wild type stains.And there were5mutation types all together.Sequencing analysis revealed that PCR products of5SSCP negative specimens had no sequence changes ......