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[Molecular characterization genetic diversity of extended-spectrum β-lactamases-harboring conjugative plasmids identified in multi-drug resistant Escherichia coli isolated from food in China].

Objective: The purpose of this study was to investigate the molecular characteristics of ESBL-encoding conjugative plasmid identified in muti-drug resistant Escherichia coli isolated from food. Methods: 465 Escherichia coli isolates were collected from national foodborne disease surveillance net from 2013 to 2014 (salad, n=159; meat, n=102; processed meat, n=95; cakes/rice, n=46; cooked dish, n=63). ESBLs strain was detected by Mueller-Hinton agar plate, and then its drug resistance was tested by agar dilution method. Polymerase chain reaction (PCR) and DNA sequencing were performed to identify the corresponding ESBL genes. Plasmids were typed by PCR-based replicon typing and their characteristics were determined by S1-nuclease pulsed-field gel electrophoresis method. Broth mating assays were carried out for all isolates to determine whether the ESBL marker could be transferred by conjugation. Results: 12 E. coli were found to be resistant to cefotaxime, and all of which were confirmed as ESBLs. The 12 isolates all carried different types of CTX-M genes resistant to drug, and 7 of which carried TEM type as well. All 12 isolates contained at least one plasmid and some had four plasmids, with size ranging from 47-to 220-kb by S1-PFGE anaylsis. Seven isolates demonstrated the ability to transfer their cefotaxime resistance marker to the recotper strain J53 by only one plasmid. Conclusion: This study highlights the diversity of the multi-drug resistant E. coli and also the diversity of ESBL genes in China. Plasmids carrying these genes poses a serious threat to food safety in China.