野生动物分离菌耐药基因多重PCR检测及序列分析  

Detection and sequence analysis of antibiotic resistance genes in strains isolated from wild animals

作　　者：张安云[1] 王红宁[2] 周万蓉[1] 柳萍[3] 黄勇[3] 杨鑫[3] 夏青青[3] 刘立[3] 

机构地区：[1]四川农业大学资源环境学院,四川雅安625014 [2]四川大学"985工程"西南资源环境与灾害防治科技创新平台,四川成都610064 [3]四川农业大学动物科技学院,四川雅安625014

出　　处：《畜牧与兽医》2007年第3期1-4,共4页Animal Husbandry & Veterinary Medicine

基　　金：国家"十五"重点科技攻关子课题(2002BA514A-12-5);国家农业科技成果转化基金项目(合同号:05EFN215100243)

摘　　要：调查来源于16种四川地区野生动物粪样的24株革兰氏阴性肠杆菌中氨基糖苷类抗生素四种修饰酶基因(am inoglycosides-mod ifying en-zym es,AMEs)的分布情况。应用K-B法进行常规药敏试验,采用多重PCR技术检测分离菌中四种AMEs(aphA3、aacC4、aadA、aacC2)基因并进行序列分析。药敏试验显示分离菌对链霉素的耐药率最高(8/24),对新霉素的耐药率最低(1/24%);多重PCR检测结果显示,aphA3、aa-dA、aacC2基因的检出率分别为:7/24、3/24和2/24,而所有菌株中均未检出aacC4基因;序列分析表明扩增产物与Genbank中的相应序列有很高的同源性(≥97.8%)。Twenty-four gram-negative strains were isolated from the feces of 16 kinds of wild animals lived in Bifeng canyon, Ya＇an, SiChuan. antibiotic resistance tests of 24 bacteria were done by K-B method, and 4 pairs of primers were designed according to the sequence of AMEs genes （ aphA3, aacCA, aadA and aacC2） published in Genbank. The drug-resistance rate for streptomycin was the highest in the isolates （8/24） , and that for neomycin was the lowest （1/24）. The results showed that three AMEs genes （aphA3, aadA, aacC2） were obtained respectively, and the detection frequency were 7/24, 3/24, 2/24. aacCA gene was not obtained. The homologous rate of each PCR product was very high compared with the sequences from Genbank （ ≥97. 8% ）.

关 键 词：野生动物 分离菌(G^-) 氨基糖苷类 耐药基因 多重PCR 

分 类 号：S858.9[农业科学—临床兽医学]