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[中国食品中分离出的多重耐药菌携带的超广谱β-内酰胺酶接合质粒的分子特征及遗传多样性]

[Molecular characterization genetic diversity of extended-spectrum β-lactamases-harboring conjugative plasmids identified in multi-drug resistant isolated from food in China].

作者信息

Bai L, Pan Z, Xu J, Li F Q

机构信息

Key Laboratory of Food Safety Risk Assessment of Health, China National Center for Food Safety Risk Assessment, Beijing 100021, China.

出版信息

Zhonghua Yu Fang Yi Xue Za Zhi. 2017 Jul 6;51(7):610-614. doi: 10.3760/cma.j.issn.0253-9624.2017.07.007.

Abstract

The purpose of this study was to investigate the molecular characteristics of ESBL-encoding conjugative plasmid identified in muti-drug resistant isolated from food. 465 isolates were collected from national foodborne disease surveillance net from 2013 to 2014 (salad, 159; meat, 102; processed meat, 95; cakes/rice, 46; cooked dish, 63). ESBLs strain was detected by Mueller-Hinton agar plate, and then its drug resistance was tested by agar dilution method. Polymerase chain reaction (PCR) and DNA sequencing were performed to identify the corresponding ESBL genes. Plasmids were typed by PCR-based replicon typing and their characteristics were determined by S1-nuclease pulsed-field gel electrophoresis method. Broth mating assays were carried out for all isolates to determine whether the ESBL marker could be transferred by conjugation. 12 were found to be resistant to cefotaxime, and all of which were confirmed as ESBLs. The 12 isolates all carried different types of CTX-M genes resistant to drug, and 7 of which carried TEM type as well. All 12 isolates contained at least one plasmid and some had four plasmids, with size ranging from 47-to 220-kb by S1-PFGE anaylsis. Seven isolates demonstrated the ability to transfer their cefotaxime resistance marker to the recotper strain J53 by only one plasmid. This study highlights the diversity of the multi-drug resistant and also the diversity of ESBL genes in China. Plasmids carrying these genes poses a serious threat to food safety in China.

摘要

本研究的目的是调查从食品中分离出的耐多药菌株中编码超广谱β-内酰胺酶(ESBL)的接合质粒的分子特征。2013年至2014年从国家食源性疾病监测网收集了465株分离株(沙拉,159株;肉类,102株;加工肉类,95株;蛋糕/米饭,46株;熟食,63株)。通过穆勒-欣顿琼脂平板检测ESBLs菌株,然后用琼脂稀释法检测其耐药性。进行聚合酶链反应(PCR)和DNA测序以鉴定相应的ESBL基因。通过基于PCR的复制子分型对质粒进行分型,并通过S1-核酸酶脉冲场凝胶电泳法确定其特征。对所有分离株进行肉汤交配试验,以确定ESBL标记是否可以通过接合转移。发现12株对头孢噻肟耐药,所有这些均被确认为ESBLs。这12株分离株均携带不同类型的耐药CTX-M基因,其中7株还携带TEM型基因。所有12株分离株均至少含有一个质粒,有些含有四个质粒,通过S1-PFGE分析,大小范围为47至220 kb。7株分离株仅通过一个质粒就能够将其头孢噻肟耐药标记转移至受体菌株J53。本研究突出了中国耐多药菌株的多样性以及ESBL基因的多样性。携带这些基因的质粒对中国食品安全构成严重威胁。

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