一种用于从培养细菌、粪便和环境样本中检测三种黏菌素抗性基因的多重SYBR Green实时荧光定量PCR检测方法。
A Multiplex SYBR Green Real-Time PCR Assay for the Detection of Three Colistin Resistance Genes from Cultured Bacteria, Feces, and Environment Samples.
作者信息
Li Jiyun, Shi Xiaomin, Yin Wenjuan, Wang Yang, Shen Zhangqi, Ding Shuangyang, Wang Shaolin
机构信息
Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University, Beijing, China.
Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety and Beijing Laboratory for Food Quality and Safety, Beijing, China.
出版信息
Front Microbiol. 2017 Oct 27;8:2078. doi: 10.3389/fmicb.2017.02078. eCollection 2017.
The aim of the study was to develop a multiplex assay for rapid detection of , and , 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 genes, and applied to cultured bacteria, feces and soil samples. All three genes could be detected with a lower limit of 10 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 -positive , which could facilitate the screening of 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.
本研究的目的是开发一种多重检测方法，用于快速检测肠杆菌科细菌中由质粒介导的对黏菌素耐药的一组基因 、 和 。已设计了一种基于SYBR Green的实时PCR检测方法来检测这些基因，并应用于培养的细菌、粪便和土壤样本。所有这三个基因均能在低至10个培养细菌的情况下被检测到。该检测方法具有高度的特异性和敏感性，且无假阳性结果。当应用于含有阳性 的粪便和土壤样本时，该检测方法也具有结论性，这不仅有助于在细菌中筛选这些基因，还能直接从环境中进行筛选。这种简单、快速、灵敏且特异的多重检测方法将有助于临床医学和畜牧业中对黏菌素耐药性的快速筛查。
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