畜牧兽医学报 ›› 2008, Vol. 39 ›› Issue (8): 1075-1080.doi:

• 预防兽医 • 上一篇    下一篇

山东地区产CMY-2型β-内酰胺酶鸡源大肠杆菌的出现及相关特性研究

张永宁,柳洪洁,牛钟相   

  1. 山东农业大学 动物医学院,泰安 271018
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-08-24 发布日期:2008-08-24
  • 通讯作者: 牛钟相

Appearance of blaCMY-2-producing Escherichia coli Isolates of Chicken Origin in Shandong Province and Studies on Their Relevant Characterizations

ZHANG Yong-ning, LIU Hong-jie, NIU Zhong-xiang   

  1. College of Veterinary Medicine, Shandong Agricultural University, Taian 271018, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-08-24 Published:2008-08-24
  • Contact: NIU Zhong-xiang

摘要: 从山东省不同地区分离到126株鸡源大肠杆菌,利用PCR对CMY-2基因进行检测,其中15株呈CMY-2阳性;核苷酸序列分析结果表明,除JN10外,其余序列与GenBank中已发表的CMY-2序列完全相同;采用K-B法检测15株大肠杆菌对抗生素的敏感性,结果发现,菌株均耐头孢噻吩、头孢唑啉、头孢克洛、头孢克肟、氨苄西林、链霉素、四环素、氯霉素和甲氧苄啶/磺胺甲噁唑,其中3株(JN5、JN9和HZ9)耐头孢曲松和头孢他啶,但所有菌株对头孢吡肟敏感;改良三维试验证实,15株大肠杆菌均产AmpC β-内酰胺酶,其中4株同时产超广谱β-内酰胺酶(ESBLs);接合试验表明,携带CMY-2基因的质粒能在大肠杆菌间传递;脉冲场凝胶电泳(PFGE)排除了所有菌株来源于同一大肠杆菌克隆的可能性。本研究为探讨和解决细菌耐药性问题提供了重要的理论依据和研究基础,同时对了解山东地区大肠杆菌的耐药状况和耐药性散播情况提供了重要的信息。

Abstract: 126 E. coli isolates of chicken origin were collected from different areas in Shandong province. With the specific primers for the blaCMY-2 gene, 15 isolates were demonstrated to be positive for blaCMY-2 by PCR amplification. Nucleotide sequence analysis demonstrated that the blaCMY-2 sequences of these isolates were identical to that published from GenBank except for strain JN10. Susceptibilities of the 15 isolates to the selected antimicrobial agents were determined by the Kirby-Bauer disk diffusion method, which demonstrated that all the 15 isolates were resistant to cefalothin, cefazolin, cefaclor, cefixime, ampicillin, streptomycin, tetracycline, chloramphenicol and trimethoprim/sulfamethoxazole. Three (JN5, JN9, and HZ9) of the 15 isolates were also resistant to ceftriaxone and ceftazidime. But all isolates were sensitive to cefepime. The modified three dimensional test demonstrated that all the 15 isolates produced CMY-2-type AmpC β-lactamases and four of the 15 isolates produced extendedspectrum βlactamases (ESBLs) at the same time. In addition, the plasmids harbouring the blaCMY-2 gene were demonstrated to be successfully transferred from E.coli isolates to the recipient E.coli DH5α (sodiumazide resistant mutation) by conjugation experiment. The possibility of the 15 isolates deriving from the same clone was excluded by pulsed-field gel electrophoresis. This study provides a very important theory and research foundation for probing into and resolving the problems of drug-resistance among bacteria. Besides, it also provides important surveillance information for knowing the drug-resistance condition of these E.coli isolates and their dissemination in Shandong province.