Multiplex PCR Identification and 2b-RAD Sequence Analysis of Porcine Diarrheagenic Escherichia coli in Different Regions in China
In order to make clear the status of contamination and phylogenetic relationship of diarrheagenic Escherichia coli(E.coli)in raw pork products in different regions of China and to reduce the public health hazards that might occur therefrom,diarrheagenic E.coli strains were identified by multiplex PCR from 283 E.coli strains isolated from the surface of pig carcasses in North China,East China,Central China and Southwest China in recent years,then 2b-RAD sequencing and gene typing were carried out. It was found that,by multiplex PCR,36 strains of diarrheagenic E.coli were detected out,with the detection rate of 12.7%,among which,the detection rate in Southwest China(26.92%)was significantly higher than those in North China(13.33%),East China(10.57%)and Central China(10.81%);for the five types of diarrhea,enterotoxigenic E.coli(ETEC),enteropathogenic E.coli(EPEC),enteroaggregative E.coli(EAEC),enterohemorrhagic E.coli(EHEC)and enteroinvasive E.coli(EIEC)accounted for 33.33%,30.56%,25.00%,5.56% and 5.56%,respectively. After 2b-RAD sequencing for species clustering analysis,it was found that the 36 strains could be classified into 8 subgroups;it was shown that,by phylogenetic tree analysis,the strains with close genetic relationship belonged to similar diarrheagenic types,and their isolation time was also close,in particular,the strains from the same region were with closely genetic relationship. It was concluded that the diarrheagenic E.coli contaminated in raw pork products mainly included three types of ETEC,EPEC and EAEC,the diarrheagenic E.coli strains were distributed with regional diversity,and their genetic relationships were with certain spatial and temporal specificity. In conclusion,some scientific data and technical support were provided to prevent and control the containment with diarrheagenic E.coli during the process of pig slaughtering and to safeguard hygiene and safety of pork products.