内蒙古中东部地区乳源性大肠埃希菌PCR检测及其耐药性分析  

PCR determination and analysis of drug resistance of Escherichia coli from milk in Inner Mongolia

作　　者：刘洋[1,2] 布日额[1,3] 吴金花[1,3] 郭闯[1,3] 锡林高娃[1,3] 刘燕[1,3] 薛晓阳[1,2] 

机构地区：[1]内蒙古民族大学生命科学学院,内蒙古通辽028043 [2]内蒙古民族大学动物科技学院,内蒙古通辽0280432 [3]内蒙古民族大学乳源性致病菌研究所,内蒙古通辽0280432

出　　处：《中国病原生物学杂志》2012年第7期503-506,共4页Journal of Pathogen Biology

摘　　要：目的分离牛乳源性大肠埃希菌(E.coli)并进行PCR检测及耐药性分析。方法根据GenBank上公布的牛源致病性大肠埃希菌基因序列,利用Primer5.0和DNAMAN生物信息学软件,根据该致病菌基因组序列16S～23SrRNA间隔区保守序列设计并合成1对引物,对牛乳中分离的大肠埃希菌进行基因扩增和序列测定,同时利用琼脂平板扩散方法分析阳性菌株的耐药状况。结果大肠埃希菌基因PCR扩增产物为396bp,与预期片段大小相符;测序结果与GenBank上公布的牛源大肠埃希菌(X80731.1)基因序列相似性为97.22%,与标准菌株大肠埃希菌(CVCC247)基因序列相似性为100%。分离菌株普遍对卡那霉素、氧氟沙星、氟哌酸高度敏感,对四环素、氨苄青霉素高度耐药。结论成功分离出牛乳源大肠埃希菌菌株,该组分离菌对四环素和氨苄青霉素高度耐药,为进一步建立牛乳中致病性大肠埃希菌的分子检测方法及指导临床用药提供了依据。Objectives To isolate and detect Escherichia coli in milk with PCR and analyze the drug resistance of E.coli.Methods A pair of primers was designed and synthesized in accordance with the 16S-23S rRNA intergenic regions of gene sequences of cattle E.coli gene sequences in GenBank using the biological software Primer 5.0 and DNAMAN.The E.coli isolated from milk was cloned and sequenced.Drug resistance was analyzed using the agar diffusion method. Results Results showed that E.coli target gene was 396 bp in length.The cloned E.coli sequence had similarity to E.coli（X80731.1） in GenBank of 97.22% and similarity to the gene sequence of the standard strain of E.coli（CVCC247） of 100%.Conclusion Bovine mastitis E.coli had been isolated.The isolated E.coli was highly resistant to tetracycline and penicillin.These results provide the basis for further establishment of molecular detection of E.coli in milk and guidance for clinical treatment.

关 键 词：大肠埃希菌 药物敏感性 序列分析 

分 类 号：R378.21[医药卫生—病原生物学]