AUTHOR=Li Jiyun , Shi Xiaomin , Yin Wenjuan , Wang Yang , Shen Zhangqi , Ding Shuangyang , Wang Shaolin 

TITLE=A Multiplex SYBR Green Real-Time PCR Assay for the Detection of Three Colistin Resistance Genes from Cultured Bacteria, Feces, and Environment Samples

JOURNAL=Frontiers in Microbiology

VOLUME=8

YEAR=2017

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2017.02078

DOI=10.3389/fmicb.2017.02078

ISSN=1664-302X

ABSTRACT=<p>The aim of the study was to develop a multiplex assay for rapid detection of <italic>mcr-1, mcr-2</italic>, and <italic>mcr-3</italic>, a group of genes of conferring resistance to colistin mediated by plasmid in Enterobacteriaceae. A SYBR Green based real-time PCR assay has been designed to detect the <italic>mcr</italic> genes, and applied to cultured bacteria, feces and soil samples. All three <italic>mcr</italic> genes could be detected with a lower limit of 10<sup>2</sup> cultured bacteria. This test was highly specific and sensitive, and generated no false-positive results. The assay was also conclusive when applied to feces and soil samples containing <italic>mcr-1</italic>-positive <italic>Escherichia coli</italic>, which could facilitate the screening of <italic>mcr</italic> genes not only in the bacteria, but also directly from the environment. This simple, rapid, sensitive, and specific multiplex assay will be useful for rapid screening of the colistin resistance in both clinical medicine and animal husbandry.</p>