| Summary: | Carbapenem-resistant <i>Enterobacterales</i> (CRE) species are top priority pathogens according to the World Health Organization. Rapid detection is necessary and useful for their surveillance and control globally. This study developed a multiplex polymerase chain reaction (mPCR) detection of the common carbapenemase genes NDM, KPC, and OXA-48-like, together with identification of <i>Escherichia coli,</i> and distinguished a <i>Klebsiella pneumoniae</i> complex to be <i>K. pneumoniae</i>, <i>K. quasipneumoniae</i>, and <i>K. variicola</i>. Of 840 target <i>Enterobacterales</i> species, 190 <i>E. coli</i>, 598 <i>K. pneumoniae</i>, 28 <i>K. quasipneumoniae</i>, and 23 <i>K. variicola</i>. with and without NDM, KPC, or OXA-48-like were correctly detected for their species and carbapenemase genes. In contrast, for the <i>Enterobacterales</i> species other than <i>E. coli</i> or <i>K. pneumoniae</i> complex with carbapenemase genes, the mPCR assay could detect only NDM, KPC, or OXA-48-like. This PCR method should be useful in clinical microbiology laboratories requiring rapid detection of CRE for epidemiological investigation and for tracking the trends of carbapenemase gene dynamics.
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