Rapid Increase in the IS26-Mediated cfr Gene in E. coli Isolates with IncP and IncX4 Plasmids and Co-Existing cfr and mcr-1 Genes in a Swine Farm.
Rapid Increase in the IS26-Mediated cfr Gene in E. coli Isolates with IncP and IncX4 Plasmids and Co-Existing cfr and mcr-1 Genes in a Swine Farm.
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IS 26介导的cfr基因在E.猪场分离到的携带IncP和IncX 4质粒并同时携带cfr和mcr-1基因的大肠杆菌
DOI：
10.3390/pathogens10010033
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发表时间：
2021-01-03
期刊：
影响因子：
--
通讯作者：
Zeng Z
中科院分区：
文献类型：
--
作者：
Ma Z;Liu J;Chen L;Liu X;Xiong W;Liu JH;Zeng Z
目的研究2014-2017年某典型猪场大肠杆菌cfr基因分离株的分子流行病学特征。用PCR扩增法从617株大肠杆菌中筛选出cfr基因。采用药敏试验、脉冲场凝胶电泳（PFGE）、S1-PFGE、southern blotting杂交和cfr基因遗传背景对所有cfr阳性大肠杆菌分离株进行分析。以大肠杆菌C600为受体菌株，45株含mcr-1-cfr的大肠杆菌为供体菌株，采用肉汤配种法进行了偶联实验。利用Illumina Miseq 2500检测到质粒pHNEP124和pHNEP129。85株（13.7%）大肠杆菌cfr基因阳性，cfr基因阳性率从2014年的1.6%显著上升至2017年的29.1%。脉冲场凝胶电泳（PFGE）分析表明，cfr基因在大肠杆菌分离株中的传播主要是水平转移。此外，cfr基因主要位于28.8 ~ 60kb大小的质粒上，并且cfr基因两侧有两个相同取向的IS26拷贝。序列分析表明，共携带cfr和mcr-1基因的质粒pHNEP124和pHNEP129分别属于IncP质粒和IncX4质粒。综上所述，本研究首次报道了cfr基因在大肠杆菌分离株中的高流行率，也首次报道了携带mcr-1和cfr基因的IncP和IncX4质粒的全基因组序列。携带cfr/mcr-1的质粒在大肠杆菌分离株中的发生和传播情况需要进一步监测。
This paper aimed to investigate the molecular epidemiological features of the cfr gene in E. coli isolates in a typical swine farm during 2014–2017. A total of 617 E. coli isolates were screened for the cfr gene using PCR amplification. A susceptibility test, pulsed-field gel electrophoresis (PFGE), S1-PFGE, southern blotting hybridization, and the genetic context of the cfr gene were all used for analyzing all cfr-positive E. coli isolates. A conjugation experiment was conducted with the broth mating method using E. coli C600 as the recipient strain and 45 mcr-1-cfr-bearing E. coli isolates as the donor strain. Plasmids pHNEP124 and pHNEP129 were revealed by Illumina Miseq 2500. Eighty-five (13.7%) E. coli isolates were positive for the cfr gene and the prevalence of the cfr gene had significantly increased from 1.6% in 2014 to 29.1% in 2017. The Pulsed-Field Gel Electrophoresis (PFGE) analysis indicated that the spread of the cfr gene among E. coli isolates was mainly due to horizontal transfer. In addition, the cfr gene was primarily located on the plasmids between 28.8-kb to 60-kb in size, and the cfr gene was flanked by two copies of IS26 with the same orientation. Sequence analysis suggested that the plasmids pHNEP124 and pHNEP129 co-harboring the cfr and mcr-1 genes belonged to the plasmids IncP plasmid and IncX4 plasmid, respectively. In conclusion, this is the first study to report the high prevalence of the cfr gene among E. coli isolates and the first report of the complete genome sequence of IncP and IncX4 plasmids carrying the mcr-1 and cfr genes. The occurrence and dissemination of the cfr/mcr-1-carrying plasmids among E. coli isolates need further surveillance.
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作者：
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