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Rapid detection of human origin colistin-resistance genes mcr-1, mcr-3, mcr-8, mcr-10 in clinical fecal samples
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Rapid detection of human origin colistin-resistance genes mcr-1, mcr-3, mcr-8, mcr-10 in clinical fecal samples
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Rapid detection of human origin colistin-resistance genes mcr-1, mcr-3, mcr-8, mcr-10 in clinical fecal samples
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Journal Article
Rapid detection of human origin colistin-resistance genes mcr-1, mcr-3, mcr-8, mcr-10 in clinical fecal samples
2021
Overview
Plasmid-mediated colistin-resistance genes have been reported in human origin clinical samples worldwide which raises its threats to human infections. Notably,
mcr-1
,
mcr-3
,
mcr-8
, and
mcr-10
have been reported isolated directly from clinical samples which creates more seriously threaten to human health than other
mcr
gene types. A multiplex polymerase chain reaction (Multi-PCR) protocol was developed to detect and genotype mobile colistin-resistance genes (
mcr-1
,
mcr-3
,
mcr-8
,
mcr-10
) in
Enterobacteria
for clinical laboratory purposes. We first designed four pairs of new primers for the amplification of
mcr
-1,
mcr
-3,
mcr
-8, and
mcr
-10 gene respectively to achieve stepwise separation of amplicons between 216 and 241 bp, and complete this Multi-PCR system with the assistance of another pair of universal primer. Among which the forward primers for
mcr-8
and
mcr-10
amplicons were identical. The protocol was validated by testing 11 clinical isolates of
Escherichia coli
and 3 clinical isolates of
Klebsiella
from human origin, each well characterized and prospectively validated. The Multi-PCR assay showed full concordance with whole-genome sequence data and displayed higher sensitivity and 100% specificity. The assay could detect all variants of the various
mcr
alleles described. The Multi-PCR assay successfully genotyped of
mcr
alleles described in one test.
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