Frontiers in Microbiology (Feb 2019)

PCR-Dipstick-Oriented Surveillance and Characterization of mcr-1- and Carbapenemase-Carrying Enterobacteriaceae in a Thai Hospital

  • Rathina Kumar Shanmugakani,
  • Rathina Kumar Shanmugakani,
  • Rathina Kumar Shanmugakani,
  • Yukihiro Akeda,
  • Yukihiro Akeda,
  • Yukihiro Akeda,
  • Yo Sugawara,
  • Warawut Laolerd,
  • Narong Chaihongsa,
  • Suntariya Sirichot,
  • Norihisa Yamamoto,
  • Norihisa Yamamoto,
  • Norihisa Yamamoto,
  • Hideharu Hagiya,
  • Hideharu Hagiya,
  • Hideharu Hagiya,
  • Daiichi Morii,
  • Daiichi Morii,
  • Yoshihiro Fujiya,
  • Yoshihiro Fujiya,
  • Yoshihiro Fujiya,
  • Isao Nishi,
  • Hisao Yoshida,
  • Hisao Yoshida,
  • Hisao Yoshida,
  • Dan Takeuchi,
  • Noriko Sakamoto,
  • Kumthorn Malathum,
  • Pitak Santanirand,
  • Kazunori Tomono,
  • Kazunori Tomono,
  • Shigeyuki Hamada

DOI
https://doi.org/10.3389/fmicb.2019.00149
Journal volume & issue
Vol. 10

Abstract

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Colistin is used as an alternative therapeutic for carbapenemase-producing Enterobacteriaceae (CPE) infections which are spreading at a very high rate due to the transfer of carbapenemase genes through mobile genetic elements. Due to the emergence of mcr-1, the plasmid-mediated colistin resistance gene, mcr-1-positive Enterobacteriaceae (MCRPEn) pose a high risk for the transfer of mcr-1-carrying plasmid to CPE, leading to a situation with no treatment alternatives for infections caused by Enterobacteriaceae possessing both mcr-1 and carbapenemase genes. Here, we report the application of PCR-dipstick-oriented surveillance strategy to control MCRPEn and CPE by conducting the PCR-dipstick technique for the detection of MCRPEn and CPE in a tertiary care hospital in Thailand and comparing its efficacy with conventional surveillance method. Our surveillance results showed a high MCRPEn (5.9%) and CPE (8.7%) carriage rate among the 219 rectal swab specimens examined. Three different CPE clones were determined by pulsed-field gel electrophoresis (PFGE) whereas only two MCRPEn isolates were found to be closely related as shown by single nucleotide polymorphism-based phylogenetic analysis. Whole genome sequencing (WGS) and plasmid analysis showed that MCRPEn carried mcr-1 in two plasmids types—IncX4 and IncI2 with ~99% identity to the previously reported mcr-1-carrying plasmids. The identification of both MCRPEn and CPE in the same specimen indicates the plausibility of plasmid-mediated transfer of mcr-1 genes leading to the emergence of colistin- and carbapenem-resistant Enterobacteriaceae. The rapidity (<2 h) and robust sensitivity (100%)/specificity (~99%) of PCR-dipstick show that this specimen-direct screening method could aid in implementing infection control measures at the earliest to control the dissemination of MCRPEn and CPE.

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