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Multidrug-resistant (MDR)  Escherichia coli  poses a great challenge for public health in recent decades. Polymyxins have been reconsidered as a valuable therapeutic option for the treatment of infections caused by MDR  E. coli.  A plasmid-encoded colistin resistance gene  mcr-1  encoding phosphoethanolamine transferase has been recently described in Enterobacteriaceae. In this study, a total of 123  E .  coli  isolates obtained from patients with diarrheal diseases in China were used for the genetic analysis of colistin resistance in clinical isolates. Antimicrobial resistance profile of polymyxin B (PB) and 11 commonly used antimicrobial agents were determined. Among the 123  E. coli  isolates, 9 isolates (7.3%) were resistant to PB and PCR screening showed that seven (5.7%) isolates carried the  mcr-1  gene. A hybrid sequencing analysis using single-molecule, real-time (SMRT) sequencing and Illumina sequencing was then performed to resolve the genomes of the seven  mcr-1  positive isolates. These seven isolates harbored multiple plasmids and are MDR, with six isolates carrying one  mcr-1  positive plasmid and one isolate (14EC033) carrying two  mcr-1  positive plasmids. These eight  mcr-1  positive plasmids belonged to the IncX4, IncI2, and IncP1 types. In addition, the  mcr-1  gene was the solo antibiotic resistance gene identified in the  mcr-1  positive plasmids, while the rest of the antibiotic resistance genes were mostly clustered into one or two plasmids. Interestingly, one  mcr-1  positive isolate (14EC047) was susceptible to PB, and we showed that the activity of MCR-1-mediated colistin resistance was not phenotypically expressed in 14EC047 host strain. Furthermore, three isolates exhibited resistance to PB but did not carry previously reported  mcr -related genes. Multilocus sequence typing (MLST) showed that these  mcr-1  positive  E .  coli  isolates belonged to five different STs, and three isolates belonged to ST301 which carried multiple virulence factors related to diarrhea. Additionally, the  mcr-1  positive isolates were all susceptible to imipenem (IMP), suggesting that IMP could be used to treat infection caused by  mcr-1  positive  E. coli  isolates. Collectively, this study showed a high occurrence of  mcr-1  positive plasmids in patients with diarrheal diseases of Guangzhou in China and the abolishment of the MCR-1 mediated colistin resistance in one  E. coli  isolate.

Antimicrobial Resistance Profile of mcr-1 Positive Clinical Isolates of Escherichia coli in China From 2013 to 2016