肺炎克雷伯菌耐氟喹诺酮机理研究.docVIP

中医眼耳鼻喉杂志编辑部1287699781@ 肺炎克雷伯氏菌耐氟喹诺酮机制研究 梁建英 黄永茂2 于晓燕3 杜元平4 （1、3、4 四川省南充市中心医院感染科 637000 2四川省泸州医学院附属医院感染科646000） 目的：了解泸州地区肺炎克雷伯氏菌对氟喹诺酮类药物耐药机制的探讨。方法：采用NCCLS推荐的K-B法测定50株肺炎克雷伯氏菌对氟喹诺酮类药物的耐药性并初步筛选出耐环丙沙星菌株；试管稀释法测定耐环丙沙星菌株对环丙沙星和左氧氟沙星的最低抑菌浓度（MIC）；并对此23株菌GyrA的基因（gyrA）进行PCR扩增和基因序列的分析比较。结果：23株耐环丙沙星肺炎克雷伯氏菌中，20株存在gyrA变异：Ser83 (TCC））The Mechanisms of Klebsiella pneumoniae’s Resistance to Fluoroquinolones Abstract: objective: To investigate the mechanisms of Klebsiella pneumoniae’s resistance to fluoroquinolones in this area. Methods: 1. The antibiotic susceptibility test of 50 Klebsiella pneumoniae to fluoroquinolones was performed by Kerby-Bauer disk diffusion method and the strains resistant to ciprofloxacin were picked up. 2.The minimal inhibitory concentrations（MICs）of twenty-three strains Klebsiella pneumoniae to Ciprofloxacin and Levofloxacin were determined by tube dilution method. MIC testing was performed by the NCCLS reference method. 3. The genes of GyrA (gyrA) in twenty-three strains Klebsiella pneumoniae resistant to ciprofloxacin were amplified by PCR and their DNA were sequencing. Results： In 23 strains klebsiella pneumoniae resistant to ciprofloxacin, 20 strains of which was mutated in gyrA: Ser83 (TCC）→Phe (TTC）、Tyr（TAC），Asp87（GAC）→Asn（AAC）、Ala （GCC）. Conclusion：The drug resistance mechanisms of klebsiella pneumonia to fluoroquinolones and the drug target of fluoroquinolones was related to the mutations of gene gyrA . The more gene sites mutated in gene gyrA , the higher klebsiella pneumoniae resistant to fluoroquinolones. Key Words：Klebsiella pneumoniae; resistance; fluoroquinolones;Gene mutation. 近年来肺炎克雷伯氏菌感染有明显增加的趋势，并成为革兰氏阴性杆菌和医院内感染的主要致病菌之一。随着抗菌药物的广泛应用，细菌的耐药性，给临床抗感染治疗对抗生素的选择增加了难度。2006年8月～2007年5月间从泸州医学院附属医院住院病例送检标本中得到肺炎克雷伯氏菌共50株（不含同一病例不同部位重复分离菌株）。经全自动细菌鉴定仪VITEK系统确认为肺炎克雷伯氏菌。标准菌株：肺炎克雷伯杆菌ATCC700603，购自国家临床检验中心。 2.材料：环丙沙星、左氧氟沙星、加替沙星纸片购自英国OXOID公司。环丙沙星标准品（效能84.9%）、左氧氟沙星标准品（效能97.3%）购自中国药品生物制品检定所。gyrA上游引物5＇-ATGAGCGACCTTGCG-3＇，下游引物5＇-CCGTGCCGTCATAGTT-3＇（457bp）由上海生工生物技术公司合成