3株猪源大肠埃希氏菌耐药基因的初步定位

doi:10.16431/j.cnki.1671-7236.2019.04.034

摘要/Abstract

摘要：

本试验旨在对临床分离的猪源大肠埃希氏菌耐药基因进行初步定位。采用常规细菌分离培养、16S rRNA PCR扩增和序列测定方法从江西省3个规模化猪场送检的子宫脓液中分离鉴定病原菌，并通过质粒提取、转化大肠埃希氏菌DH5α感受态细胞及药敏试验对临床分离株的耐药基因进行初步定位。结果显示，分离鉴定到3株大肠埃希氏菌，其中JX-22分离株仅对氧氟沙星、大观霉素敏感，JX-26分离株仅对链霉素、氧氟沙星等4种药物敏感，JX-28分离株仅对氧氟沙星等3种药物敏感，均为多重耐药菌；3株大肠埃希氏菌均可纯化到分子质量大小不一的质粒。分离株、质粒转化菌及大肠埃希氏菌DH5α感受态细胞药敏试验对比结果显示，3株大肠埃希氏菌的耐链霉素、林可霉素、甲硝唑、氨苄西林、阿莫西林、大观霉素、丁胺卡那基因，JX-22和JX-26分离株的耐多西环素、氟苯尼考和复方新诺明基因，JX-22分离株的耐头孢曲松基因，JX-28分离株的耐头孢曲松、头孢噻肟、诺氟沙星基因均定位于细菌质粒上；JX-28分离株的耐多西环素、氟苯尼考和复方新诺明基因，JX-22分离株的耐诺氟沙星基因和JX-26分离株的耐头孢曲松、头孢噻肟基因均定位于其染色体上；3株分离株均无氧氟沙星耐药基因。本试验初步确定3株多重耐药猪源大肠埃希氏菌的大部分耐药基因定位于质粒上，为进一步研究猪源大肠埃希氏菌的耐药机理和有效控制措施奠定基础。

关键词: 大肠埃希氏菌; 多重耐药; 耐药基因; 药敏试验

Abstract:

This trial was aimed to preliminarily locate clinically isolated Escherichia coli (E.coli) resistant genes in pigs.Using conventional bacterial isolation and culture,16S rRNA PCR amplification and sequence analysis,the pathogens were isolated and identified from the uterus pus sent from three large-scale pig farms in Jiangxi province,the drug resistance genes of clinical isolates were preliminarily located by plasmid extraction,transformation of E.coli DH5α competent cells and drug susceptibility test.The results showed that 3 strains of E.coli were isolated and identified,among which JX-22 was only sensitive to ofloxacin and spectinomycin,and JX-26 was only sensitive to four drugs,such as streptomycin and ofloxacin,JX-28 was only sensitive to three drugs such as ofloxacin,all were multi-drug resistant bacteria.Three strains of E.coli could be purified to obtain plasmids of different molecular masses.The results of drug sensitivity test of isolates and plasmid transformants and E.coli DH5α competent cells showed that resistance genes of three strains of E.coli to streptomycin,lincomycin,metronidazole,ampicillin,amoxicillin,spectinomycin and amikacin,JX-22 and JX-26 isolates to doxycycline,florfenicol and cotrimoxazole,JX-22 isolate to ceftriaxone,and ceftriaxone,cefotaxime and norfloxacin genes of JX-28 isolate were located on a bacterial plasmid.Resistant to doxycycline,florfenicol and cotrimoxazole genes of JX-28 isolate,norfloxacin gene of JX-22 isolate and ceftriaxone and cefotaxime genes of JX-26 isolate were all located on their chromosomes.None of the three isolates had the ofloxacin resistance gene.In this experiment,most of the multidrug resistance genes of three multi-drug resistant E.coli were preliminarily located on the plasmid,which laid a foundation for further study on the drug resistance mechanism and effective control measures of E.coli.

Key words: E.coli; mult-drug resistance; drug resistance gene; drug sensitive test

中图分类号:

S852.61

杨行, 殷贵虎, 张文波, 唐云云. 3株猪源大肠埃希氏菌耐药基因的初步定位[J]. 《中国畜牧兽医》, 2019, 46(4): 1244-1251.

YANG Xing, YIN Guihu, ZHANG Wenbo, TANG Yunyun. Preliminary Location of Resistant Genes of Three Escherichia coli Strains from Swine[J]. , 2019, 46(4): 1244-1251.

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