浙江农业学报 ›› 2017, Vol. 29 ›› Issue (4): 534-541.DOI: 10.3969/j.issn.1004-1524.2017.04.03

• 动物科学 • 上一篇    下一篇

一株牛源肺炎克雷伯氏菌的分离鉴定与耐药基因型检测

蒙正群1, 冷依伊1, 任梅渗1, 刘亚东1, 王印1, 2, 姚学萍1, 杨泽晓1, *   

  1. 1.四川农业大学 动物医学院,四川 温江 611130;
    2.动物疫病与人类健康四川省重点实验室,四川 温江 611130
  • 收稿日期:2016-10-26 出版日期:2017-04-20 发布日期:2017-04-27
  • 通讯作者: 杨泽晓,E-mail:yzxyang2003@126.com
  • 作者简介:蒙正群(1992—),女,贵州瓮安人,硕士研究生,研究方向为预防兽医学。E-mail:m15983544718_1@163.com
  • 基金资助:
    “十二五”国家科技支撑计划(2013BAD12B04); 四川农业大学学科建设双支计划(03571806)

Isolation, identification and drug resistance detection of a calf Klebsiella pneumoniae strain

MENG Zhengqun1, LENG Yiyi1, REN Meishen1, LIU Yadong1, WANG Yin1, 2, YAO Xueping1, YANG Zexiao1, *   

  1. 1. College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang 611130, China;
    2. Key Laboratory of Animal Disease and Human Health of Sichuan Province,Wenjiang 611130, China
  • Received:2016-10-26 Online:2017-04-20 Published:2017-04-27

摘要: 从病死犊牛肺脏组织中分离到一株革兰阴性短杆菌,进行了细菌分离纯化、生化试验、16SrDNA测序鉴定、小鼠致病性试验、药敏试验,及碳青霉烯类酶与产超广谱β-内酰胺酶(ESBLs)等7种耐药基因的PCR检测。结果鉴定该菌为肺炎克雷伯氏菌,其16S rDNA扩增序列与GenBank中肺炎克雷伯氏菌同源性达99%。对小鼠致病性强。该菌分离株对氨苄西林、头孢呋辛、复方新诺明、红霉素耐药,而对碳青霉烯类、第四代头孢等受试药物均为敏感。耐药基因PCR检测仅扩增出约862 bp的SHV基因片段,与公布的SHV-1基因序列同源性达99.3%。该牛源克雷伯氏菌分离株耐药表型与耐药基因检测结果一致。

关键词: 肺炎克雷伯氏菌, 分离鉴定, 耐药基因

Abstract: A gram-negative bacillus was isolated and purified from the lung of a died calf, and was identified by morphological and cultured characteristics, biochemical tests, 16S rDNA gene sequencing, pathogenicity tests, and drug sensitivity tests. Then its drug-resistance genes were detected and analyzed by PCR methods using 7 pairs of primers for drug-resistance genes that produce carbapenemase and extended-spectrum β-lactamase (ESBLs) etc. The results showed that this isolate was identified as cow-derived Klebsiella pneumoniae, which was pathogenic to mice. Its 16S rDNA gene sequence shared 99% homology with Klebsiella pneumoniae gene sequences published in GenBank. The isolate was highly resistant with penbritin, cefuroxime, co-trimoxazole and erythromycin, while sensitive to the gross antibiotics such as carbapenems and the fourth generation cephalosporin. A resistance gene fragment about 862 bp in the length of SHV gene was amplified by PCR, which had a 99.3% homology with published SHV-1 gene sequences in GenBank. The cow-derived Klebsiella pneumoniae strain had strong virulence and drug-resistance, and its drug-resistant phenotype was consistent with the drug-resistant testing results.

Key words: Klebsiella pneumoniae, isolation and identification, drug resistance genes

中图分类号: