养殖动物及人分离大肠埃希菌染色体和质粒介导氟喹诺酮耐药机制的研究

目的 探讨从养殖动物及周围人群分离的大肠埃希菌染色体和质粒介导氟喹诺酮耐药机制. 方法 纸片扩散法和肉汤稀释法检测氟喹诺酮抗菌药物及其他抗生素的耐药性表型.PCR扩增DNA解旋酶(gyrA和gyrB)和拓扑异构酶IV(parC和parE)基因的喹诺酮耐药决定区、导致喹诺酮类抗生素耐药质粒的部分基因(qnr)以及氨基糖苷类抗生素乙酰转移酶Ib亚型cr变异体编码基因aac(6')-I b-or],PCR产物进行直接测序.接合试验确定aac(6')-I b-cr酶的可转移性以及在氟喹诺酮耐药中的作用. 结果 鸡来源的大肠埃希菌对常用抗生素的耐药率明显高于猪和周围人群来源菌株.在PCR检测的64株大肠埃希菌中,环丙沙星MIC值大于1μg/ml以上的53株均存在gyrA和/或/parC基因上出现两个位点突变和氨基酸替代,环丙沙星的MIC>16μg/ml的菌株parE基因也发生了点突变及相应氨基酸替代.未发现gyrB亚单位有氨基酸替代.鸡来源28株菌和猪来源9株菌中分别有7株(25.O%)和1株(11.1%)携带有aac(6')-I b-cr基因;aac(6')-I b-cr基因可使环丙沙星、诺氟沙星乙酰化而降低药物抗菌活性. 结论 gyrA、parC和parE碱基突变导致氨基酸置换的数量与菌株对氟喹诺酮类耐药水平呈正相关,携带aac(6')-I b-cr基因的质粒在细菌氟喹诺酮耐药上也具有一定作用.

Chromosome-and plasmid-mediated fluoroquinolones-resistance in Escherichia coli strains isolated from food animals and healthy people around farm

Objective To study on chromosome-and plasmid-mediated fluoroquinolones-resistant in Escherichia coli isolated from fecal samples of chicken,swine and people around the farm.Methods Anti-microbial susceptibility testing was carried out by disk diffusion testing and bmth microdilution testing.gyrA,gyrB,parC,pareE,qnr and aac(6')-I b-cr were examined by PCR,and the products were sequenced.Ex-presion of aac(6')-I b-cr by conjunction was tested too.Results The resistance to antimicmbial agents was much higher in strains isolated from chicken than that from swine and human.Among the E coli strains examined by PCR,most resistant strains carried two mutations in gyrA and/or two mutations in parC.In ad-dition,some resistant strains had mutations in parE with MIC of ciprofloxacin>16μg/ml.No(resistance) mutation was found in gyrB.Seven strains(25.O%)and one strain(11.1%)had aac(6)-I b-cr,variant isolated from chicken and swine,respectively.The strains harboring cr variant enzyme reduced the suscepti-bility to ciprofloxacin and norfloxacin by N-acetylation of the drugs. Conclusion There is a close relation-ship between high level quinolone resistance and the numbers of amino acid exchange in DNA gyrase and to-poisomeraae IV,and aac(6)-I b-cr may play some role for fluoroquinolone resistance.