羊致病性大肠杆菌的分离鉴定、毒力基因及耐药基因分析

Isolation,Identification,Virulence Gene and Drug Resistance Analysis of Pathogenic Escherichia Coli from Sheep

为了确定某羊场呼吸道症状死亡羊的病因,试验通过对病死羊肺脏等病料进行了细菌分离培养和纯化、动物感染试验、16S rDNA PCR检测和序列分析鉴定;并应用PCR方法对I型菌毛(fimH)、P型菌毛(papC)、α溶血素(hlyA)、毒力岛(eaeA)、外膜蛋白A(ompA)、致病岛(fyuA)、黏附素相关基因(mat、K99)、耶尔森强毒力岛(irp2)、温度敏感性血细胞凝集素(TSH)等毒力基因进行检测,对氨基糖苷类(aadA1)、喹诺酮类(parC)、四环素类(sul1)、磺胺类(sul2)、氯霉素类(floR、gyrA)、β-内酰胺类(tetA、tetB)等耐药基因进行检测。结果表明该分离株为致病性羊大肠杆菌,分离株携带fimH、ompA、papC、mat四种毒力基因和aadA1、gyrA、parC、sul1、floR 5种耐药基因。研究为有效预防和控制羊大肠杆菌病奠定了基础。

In order to determine the cause of the dead sheep with respiratory symptoms in a sheep farm,the experiment was carried out including bacterial isolation,culture and purification,animal infection test,16S rDNA PCR detection and sequence analysis and identification of the diseased materials such as the lungs of the dead sheep.The fimH,papC,fimbriae were detected by PCRαHemolysin(hlyA),virulence Island(eaeA),outer membrane protein A(ompA),pathogenicity island(fyuA),adhesin related genes(mat,K99),yersen virulence Island(irp2),temperature sensitive hemagglutinin(tsh)and other virulence genes were detected Chloramphenicols(floR,gyrA)β-Lactams(tetA,tetB)resistance genes were detected.The results showed that the isolate was confirmed to be pathogenic Escherichia coli,and it carried four virulence genes of fimH,ompA,papC and mat,and five drug resistance genes of aada1,gyrA,parC,sul1 and floR.This study laid a foundation for effective prevention and control of sheep Escherichia coli disease.