Metallo-β-lactamases (MBL) producing Pseudomonas aeruginosa isolates have been well characterized. Quinolones are commonly used in the treatment of carbapenem-resistant P. aeruginosa infections; however, data about PMQR in this species are scarce. The objective of this study was to report the simultaneous presence of qnrS and blaVIM-11 in P. aeruginosa, and to characterize the qnrS-harboring plasmid. Thirty-eight carbapenem-resistant P. aeruginosa isolates were recovered from a hospital in Buenos Aires during 2012. Screening for MBL was assessed by the double disk synergy test using EDTA and carbapenem discs. Plasmid DNA extraction was performed by a method using phenol-chloroform. PCR followed by sequencing was carried out to determine each MBL and PMQR allele. PCR-BseGI-RFLP was performed to detect aac-(6')-Ib-cr. The gyrA-QRDR was sequenced in those PMQR-harboring isolates. Plasmid incompatibility groups and addiction systems were characterized by PCR. The PMQR-carrying plasmid was sequenced using Illumina technology, annotated using RAST and manually curated. Eleven/38 isolates were VIM producers (blaVIM-2 and blaVIM-11) while 1/38 harbored blaIMP-13. One isolate harbored blaVIM-11 and the PMQR qnrS1; however, both markers were located in different plasmids. The qnrS1-harboring plasmid (pP6qnrS1) was 117945bp in size, presented 154 CDS and corresponded to the IncR group. In addition to qnrS1, it harbored several aminoglycoside resistance markers. Although pP6qnrS1 was non-conjugative, it presented an oriT which made it possible for this plasmid to be transferable. This is the first report on P. aeruginosa carrying both blaVIM-11 and qnrS1, plus the first detection of an IncR plasmid in Argentina.

译文

产生铜绿假单胞菌的金属-β-内酰胺酶 (MBL) 已得到很好的表征。喹诺酮类药物通常用于治疗耐碳青霉烯类药物的铜绿假单胞菌感染; 但是,该物种中有关PMQR的数据很少。这项研究的目的是报告铜绿假单胞菌中qnrS和blaVIM-11的同时存在,并表征携带qnrS的质粒。2012年从布宜诺斯艾利斯的一家医院回收了38株耐碳青霉烯的铜绿假单胞菌。通过使用EDTA和碳青霉烯类圆盘的双圆盘协同试验评估MBL的筛选。通过使用苯酚-氯仿的方法进行质粒DNA提取。进行PCR,然后进行测序以确定每个MBL和PMQR等位基因。进行PCR-BseGI-RFLP检测aac-(6 ')-Ib-cr。gyrA-QRDR在那些携带PMQR的分离株中进行了测序。通过PCR表征质粒不相容组和成瘾系统。携带PMQR的质粒使用Illumina技术进行测序,使用RAST注释并手动整理。11/38个分离株是VIM生产者 (blaVIM-2和blaVIM-11),而1/38个具有blaIMP-13。一个分离物携带blaVIM-11和PMQR qnrS1; 然而,两个标记都位于不同的质粒中。qnrS1-harboring质粒 (pP6qnrS1) 大小为117945bp,呈现154个CDS,对应于IncR组。除qnrS1外,它还具有几种氨基糖苷类抗性标记。尽管pP6qnrS1是非共轭的,但它呈现了oriT,使该质粒可以转移。这是关于同时携带blaVIM-11和qnrS1的铜绿假单胞菌的首次报道,以及在阿根廷首次检测到IncR质粒。

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