Wang Yang, Lv Yuan, Cai Jiachang, Schwarz Stefan, Cui Lanqing, Hu Zhidong, Zhang Rong, Li Jun, Zhao Qin, He Tao, Wang Dacheng, Wang Zheng, Shen Yingbo, Li Yun, Feßler Andrea T, Wu Congming, Yu Hao, Deng Xuming, Xia Xi, Shen Jianzhong
Department of Veterinary Pharmacology, College of Veterinary Medicine, China Agricultural University, Beijing, China.
Institute of Clinical Pharmacology, Peking University First Hospital, Beijing, China.
J Antimicrob Chemother. 2015 Aug;70(8):2182-90. doi: 10.1093/jac/dkv116. Epub 2015 May 14.
The oxazolidinone-resistant Enterococcus faecalis E349 from a human patient tested negative for the cfr gene and 23S rRNA mutations. Here we report the identification of a novel oxazolidinone resistance gene, optrA, and a first investigation of the extent to which this gene was present in E. faecalis and Enterococcus faecium from humans and food-producing animals.
The resistance gene optrA was identified by whole-plasmid sequencing and subsequent cloning and expression in a susceptible Enterococcus host. Transformation and conjugation assays served to investigate the transferability of optrA. All optrA-positive E. faecalis and E. faecium isolates of human and animal origin were analysed for their MICs and their genotype, as well as the location of optrA.
The novel plasmid-borne ABC transporter gene optrA from E. faecalis E349 conferred combined resistance or elevated MICs (when no clinical breakpoints were available) to oxazolidinones (linezolid and tedizolid) and phenicols (chloramphenicol and florfenicol). The corresponding conjugative plasmid pE349, on which optrA was located, had a size of 36 331 bp and also carried the phenicol exporter gene fexA. The optrA gene was functionally expressed in E. faecalis, E. faecium and Staphylococcus aureus. It was detected more frequently in E. faecalis and E. faecium from food-producing animals (20.3% and 5.7%, respectively) than from humans (4.2% and 0.6%, respectively).
Enterococci with elevated MICs of linezolid and tedizolid should be tested not only for 23S rRNA mutations and the gene cfr, but also for the novel resistance gene optrA.
从一名人类患者身上分离出的对恶唑烷酮耐药的粪肠球菌E349,其cfr基因和23S rRNA突变检测均为阴性。在此,我们报告鉴定出一种新型恶唑烷酮耐药基因optrA,并首次调查该基因在来自人类和食用动物的粪肠球菌和屎肠球菌中的存在程度。
通过全质粒测序以及随后在敏感肠球菌宿主中的克隆和表达来鉴定耐药基因optrA。转化和接合试验用于研究optrA的可转移性。对所有来自人类和动物的optrA阳性粪肠球菌和屎肠球菌分离株进行了最低抑菌浓度(MIC)、基因型以及optrA定位分析。
来自粪肠球菌E349的新型质粒携带的ABC转运蛋白基因optrA赋予了对恶唑烷酮类(利奈唑胺和替地唑胺)和酚类(氯霉素和氟苯尼考)联合耐药或MIC升高(当无临床断点时)。optrA所在的相应接合性质粒pE349大小为36 331 bp,还携带酚类输出基因fexA。optrA基因在粪肠球菌、屎肠球菌和金黄色葡萄球菌中功能性表达。在来自食用动物的粪肠球菌和屎肠球菌中检测到该基因的频率更高(分别为20.3%和5.7%),而在来自人类的菌株中检测到的频率较低(分别为4.2%和0.6%)。
对于利奈唑胺和替地唑胺MIC升高的肠球菌,不仅应检测23S rRNA突变和cfr基因,还应检测新型耐药基因optrA。