外排系统与膜通透性的改变对淋球菌高水平多重耐药的影响_临床医学论文.docVIP

外排系统与膜通透性的改变对淋球菌高水平多重耐药的影响_临床医学论文 外排系统与膜通透性的改变对淋球菌高水平多重耐药的影响_临床医学论文 作者：王冬梅，王勇，夏忠弟，田峰，邹明祥 【摘要】 　　目的 探讨外排系统与膜通透性的改变对淋球菌多重耐药的影响。方法 PCR扩增淋球菌mtrR启动子区域包含13bp回文序列在内的157个碱基片段并进行DNA测序分析；提取外膜蛋白，利用SDS进行组成型的分析。结果 5株敏感株的mtrR启动子区域未发生突变，3株高水平多重耐药株的mtrR启动子区域13bp回文序列均发生了单个A/T碱基的缺失，且均存在外膜孔蛋白表达的缺失。结论 mtrR启动子区域13bp回文序列单个A/T碱基的缺失引起的mtrCDE外排泵表达增加与外膜孔蛋白表达的缺失或下降导致的膜通透性降低，在介导淋球菌产生高水平多重耐药中起一定的作用。 【关键词】 淋球菌 多重耐药 mtr 孔蛋白 　　ABSTRACT: Objective To investigate the impact of functional enhancement of efflux pump system and reduced permeability of outer membrane on highlevel multiple resistant Neisseria gonorrhoeae. Methods Several highlevel multiple resistant isolates with erythromycin MIC=128.0mg/L, accompanied by concurrent resistance to several antimicrobial agents, were selected. 13bp inverted sequence positioned within the mtrR promoter region were amplified by PCR and directly sequenced to detect the possible gene change. The outer membrane proteins of the strains were extracted to analyze the constitutive profiles by SDSPAGE. Results There were no gene mutations in 5 sensitive strains. All the 3 highlevel multiple resistant strains contained the same mutation and exhibited a single A/T base pair deletion in 13bp inverted sequence positioned within the mtrR promoter region. Meanwhile porin protein 31ku deficiency was found in all the 3 resistant strains. Conclusion The functional enhancement of efflux pump system induced by a single A/T base pair deletion in 13bp inverted sequence positioned within the mtrR promoter region and the decreased cell envelope permeability induced by the absence of porin protein may have some effect on mediating highlevel multiple resistance in Neisseria gonorrhoeae. 　　KEY WORDS: N.gonorrhoeae; multiple 　　近年来，淋球菌多重耐药现象日益严重，且呈现出由低水平向高水平耐药发展的趋势。为探讨淋球菌高水平多重耐药的机制，本实验从58株临床菌株中筛选出了3株对红霉素高度耐药且同时耐多种药物(红霉素、氧氟沙星、诺氟沙星、壮观霉素、头孢美唑)的菌株，针对这3株高水平多重耐药株，分别对其多重耐药调节基因mtrR启动子区域和其外膜蛋白进行了研究。 　　1 材料与方法 　　1.1 菌种 　　野生型淋球菌标准株ATCC19424为香港中文大学医学院微生物系惠赠。58株临床分离菌来自中南大学湘雅医院检验科，其中5株(Ng06、Ng0