PCR-dipstick-oriented surveillance and characterization of mcr-1- and carbapenemase-carrying Enterobacteriaceae in a Thai hospital

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

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1 Citation (Scopus)

Abstract

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.

Original languageEnglish
Article number149
JournalFrontiers in Microbiology
Volume10
Issue numberFEB
DOIs
Publication statusPublished - Jan 1 2019
Externally publishedYes

Keywords

  • Carbapenemase
  • Enterobacteriaceae
  • Mcr-1
  • PCR-dipstick
  • Rapid detection

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

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  • Cite this

    Shanmugakani, R. K., Akeda, Y., Sugawara, Y., Laolerd, W., Chaihongsa, N., Sirichot, S., Yamamoto, N., Hagiya, H., Morii, D., Fujiya, Y., Nishi, I., Yoshida, H., Takeuchi, D., Sakamoto, N., Malathum, K., Santanirand, P., Tomono, K., & Hamada, S. (2019). PCR-dipstick-oriented surveillance and characterization of mcr-1- and carbapenemase-carrying Enterobacteriaceae in a Thai hospital. Frontiers in Microbiology, 10(FEB), [149]. https://doi.org/10.3389/fmicb.2019.00149