College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, China.
Animal Husbandry and Fishery Bureau, Nankang District, Ganzhou, China.
J Antimicrob Chemother. 2020 May 1;75(5):1159-1164. doi: 10.1093/jac/dkaa012.
To characterize a novel tigecycline resistance gene, tet(X6), and a novel SXT-related integrative and conjugative element (ICE), ICEPgs6Chn1, found in a tigecycline-resistant Proteus genomospecies 6 strain, T60.
Strain T60 was identified by the VITEK 2 system, biochemical reactions and an SNP-based approach. The genetic profile of strain T60 was determined by WGS analysis. ICEPgs6Chn1 was analysed by PCR, conjugation experiments and bioinformatics tools. tet(X6) was characterized by cloning and protein structure prediction.
Strain T60 was resistant to ampicillin, tetracycline, tigecycline, florfenicol, colistin and kanamycin, but susceptible to cefotaxime; it also exhibited high MICs of eravacycline (32 mg/L) and omadacycline (>64 mg/L). Only one chromosome was identified and tet(X6) was located in chromosomal ICEPgs6Chn1, a member of the SXT/R391 ICE family, of 114 368 bp and encoding the antimicrobial resistance genes floR, strB, strA, aph(3')-Ia, aac(3)-IV, aph(4)-Ia, tet(X6) and sul2. The circular intermediate of ICEPgs6Chn1 was detected by PCR and sequencing, but conjugation experiments showed that it was not self-transmissible. Cloning of the novel gene tet(X6) and protein structure prediction revealed that Tet(X6) confers tigecycline resistance.
To our knowledge, this is the first report of a novel SXT/R391 ICE in a Proteus genomospecies 6 strain. Importantly, a novel high-level tigecycline resistance gene, tet(X6), emerged for the first time in the SXT/R391 element of Proteus genomospecies 6, revealing that ICEs may serve as an important platform for the accumulation of antibiotic resistance genes.
描述一种新型替加环素耐药基因 tet(X6) 和一种新型 SXT 相关整合子-转座子元件 (ICE),ICEPgs6Chn1,该基因存在于一株对替加环素耐药的变形杆菌基因组种 6 型菌株 T60 中。
采用 VITEK 2 系统、生化反应和 SNP 方法鉴定菌株 T60。通过 WGS 分析确定菌株 T60 的遗传特征。通过 PCR、接合实验和生物信息学工具分析 ICEPgs6Chn1。通过克隆和蛋白质结构预测来研究 tet(X6)。
T60 株对氨苄西林、四环素、替加环素、氟苯尼考、黏菌素和卡那霉素耐药,但对头孢噻肟敏感;它还对依拉环素(32mg/L)和奥马环素(>64mg/L)表现出较高的 MIC 值。仅鉴定出一条染色体,tet(X6)位于染色体 ICEPgs6Chn1 中,该元件属于 SXT/R391 ICE 家族,大小为 114368bp,编码抗菌药物耐药基因 floR、strB、strA、aph(3')-Ia、aac(3)-IV、aph(4)-Ia、tet(X6)和 sul2。通过 PCR 和测序检测到 ICEPgs6Chn1 的环状中间体,但接合实验表明它不能自我转移。克隆新型基因 tet(X6)并预测蛋白质结构表明,Tet(X6)赋予替加环素耐药性。
据我们所知,这是首次在变形杆菌基因组种 6 型菌株中报道新型 SXT/R391 ICE。重要的是,一种新型高水平替加环素耐药基因 tet(X6)首次出现在变形杆菌基因组种 6 型的 SXT/R391 元件中,这表明 ICE 可能是抗生素耐药基因积累的重要平台。