National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Original Bacteria, South China Agricultural University, Guangzhou, China; Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation, South China Agricultural University, Guangzhou, China.
Department of Medical Microbiology and Parasitology, Zhejiang University School of Medicine, Hangzhou, China.
Int J Antimicrob Agents. 2018 Jun;51(6):842-847. doi: 10.1016/j.ijantimicag.2018.01.007. Epub 2018 Jan 31.
Two colistin-resistant Escherichia coli strains (FS13Z2S and FS3Z6C) possessing chromosomally encoded mcr-1 isolated from swine were characterised. Whole-genome sequencing revealed that in strain FS13Z2S mcr-1 occurred in triplicate in the chromosome with another copy encoded on a pHNSHP45-2-like IncHI2 plasmid, whereas in strain FS3Z6C only one copy mcr-1 was inserted in the chromosome. It seems likely that the triplication of chromosomal copies of mcr-1 in FS13Z2S is due to intramolecular transposition events via a composite transposon containing an mcr-1 cassette bracketed by two copies of insertion sequence ISApl1, and the pap2 gene at the insertion site was truncated by an IS1294-like element. In plasmid pFS13Z2S and the chromosome of strain FS3Z6C, only a single copy of ISApl1 was present upstream of the mcr-1 cassette. The two strains exhibited similar colistin minimum inhibitory concentrations (MICs) and featured phosphoethanolamine addition to lipid A, without regard to the copy number of mcr-1. The mcr-1-harbouring plasmid was unstable in wild-type strain FS13Z2S and was quickly lost after 7 days of passage on colistin-free Luria-Bertani broth containing 0.5% SDS, but the mcr-1 copies on the chromosome persisted. These results reveal that the single copy of mcr-1 could result in modification of lipopolysaccharide (LPS) and cause colistin resistance in E. coli. Acquisition of multiple copies of mcr-1, especially on the chromosome, would facilitate stable persistence of colistin resistance in the host strain.
从猪身上分离出的两株携带染色体编码 mcr-1 的耐药大肠杆菌(FS13Z2S 和 FS3Z6C)菌株进行了特征描述。全基因组测序显示,在 FS13Z2S 菌株中,mcr-1 以三联体形式存在于染色体中,另一个拷贝则位于 pHNSHP45-2 样 IncHI2 质粒上,而在 FS3Z6C 菌株中,mcr-1 仅插入染色体的一个拷贝中。FS13Z2S 染色体上 mcr-1 拷贝的三联体似乎很可能是由于通过含有 mcr-1 盒的复合转座子的分子内转座事件导致的,该转座子两端是两个插入序列 ISApl1,而插入位点的 pap2 基因被一个类似 IS1294 的元件截断。在质粒 pFS13Z2S 和菌株 FS3Z6C 的染色体中,mcr-1 盒上游仅存在一个 ISApl1 拷贝。这两株菌表现出相似的多粘菌素最小抑菌浓度(MIC),并在脂质 A 上添加了磷酸乙醇胺,而与 mcr-1 的拷贝数无关。mcr-1 携带的质粒在野生型菌株 FS13Z2S 中不稳定,在不含多粘菌素的 Luria-Bertani 肉汤中传代 7 天后迅速丢失,但染色体上的 mcr-1 拷贝仍然存在。这些结果表明,单个 mcr-1 拷贝可导致脂多糖(LPS)的修饰,并导致大肠杆菌对多粘菌素产生耐药性。获得多个 mcr-1 拷贝,特别是在染色体上,将有助于宿主菌株中多粘菌素耐药性的稳定持续存在。
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