新疆牛源大肠杆菌O157∶H7的分离鉴定及ERIC-PCR基因分型研究

doi:10.16431/j.cnki.1671-7236.2020.01.022

Abstract

Abstract: In order to understand the pollution status and genetic diversity of bovine Escherichia coli O157:H7 in Xinjiang,to explore the genetic relationship of isolates from different areas,and to provide experimental basis for controlling the transmission of bovine E.coli O157:H7 in cattle,in this experiment,the samples were added to EC broth (37 ℃,180 r/min),then inoculated on SMAC plate and cultured overnight in 37 ℃ incubator for about 18 h.Then,white or colorless single colonies on SMAC were inoculated the MUG medium,and cultured at 37 ℃ for 18 h.The non-fluorescent samples were inoculated on the SMAC plate,and cultured at 37 ℃ for about 18 h.The white or colorless single colonies were selected the next day for PCR identification.The positive strains with rfbE and fliC gene bands were identified.ERIC-PCR fingerprinting was used to analyze the homology among the positive strains.ERIC-PCR results showed that there were 3 groups of strains with 100% similarity.The strains isolated from the Yili region were the most diverse,with 6 types,followed by Urumqi with 4 types.The most diverse source of strains was in cluster D.It could be traced by ERIC-PCR typing.ERIC-PCR was able to distinguish the isolates of specific sampling points or species.It could demonstrate that there were some similar ERIC characteristics among strains from different sources and they were clustered in the same cluster.The E.coli O157:H7 strain selected in this study had extensive genetic diversity,which was very sensitive to detect bacterial differences among different species.Therefore,ERIC-PCR could be used as an effective tool for routine monitoring and identification of E.coli O157:H7.

Key words: cattle; E.coli O157:H7; ERIC-PCR; genetic relationship; genotyping

CLC Number:

S852.61

SU Zhanqiang, ZHANG Ling, LIU Luyao, WANG Dong, MA Xiaoyu, AIKEDAI·TULUHONG, ZHANG Yi, TONG Panpan. Isolation,Identification and ERIC-PCR Genotyping of E.coli O157:H7 from Xinjiang Cattle[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(1): 182-189.

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Isolation,Identification and ERIC-PCR Genotyping of E.coli O157:H7 from Xinjiang Cattle

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