Detection of plasmid-mediated colistin-resistant and carbapenem-resistant genes by multiplex PCR

Rujirat Hatrongjit; Anusak Kerdsin; Yukihiro Akeda; Shigeyuki Hamada

doi:10.1016/j.mex.2018.05.016

Detection of plasmid-mediated colistin-resistant and carbapenem-resistant genes by multiplex PCR

MethodsX. 2018 May 25:5:532-536. doi: 10.1016/j.mex.2018.05.016. eCollection 2018.

Authors

Rujirat Hatrongjit¹, Anusak Kerdsin², Yukihiro Akeda³, Shigeyuki Hamada⁴

Affiliations

¹ Faculty of Science and Engineering, Kasetsart University, Chalermphrakiat Sakon Nakhon Province Campus, Sakon Nakhon, Thailand.
² Faculty of Public Health, Kasetsart University, Chalermphrakiat Sakon Nakhon Province Campus, Sakon Nakhon, Thailand.
³ Department of Infection Control and Prevention, Graduate School of Medicine, Osaka University, Osaka, Japan.
⁴ Thailand-Japan Research Collaboration Center on Emerging and Re-emerging Infections, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan.

Abstract

A multiplex PCR was described to simultaneously detect mcr-1 and frequently occurring carbapenem-resistant genes including bla_KPC, bla_NDM, bla_IMP, and bla_OXA-48-like in a single reaction. The PCR product sizes of these 4 carbapenem-resistant genes were 232 bp, 438 bp, 621 bp, and 798 bp for bla_IMP, bla_OXA-48-like, bla_NDM, and bla_KPC, respectively, whereas mcr-1 revealed 1126 bp of PCR product. This protocol accurately detected those resistant genes in agreement with the reference strains, 127 local carbapenem-resistant Enterobacteriaceae, 8 mcr-1 carrying Enterobacteriaceae, and 62 carbapenem-susceptible Enterobacteriaceae. This method will be useful for laboratory application and surveillance of carbapenem and/or colistin-resistant bacteria.

Keywords: PCR; blaIMP; blaKPC; blaNDM; blaOXA-48-like; mPCR for mcr-1 and carbapenem-resistant genes; mcr-1.