Antibiotic Resistance Monitoring and Reference Laboratory, Microbiology Services-Colindale, Health Protection Agency, 61 Colindale Avenue, London NW9 5EQ, UK.
J Antimicrob Chemother. 2011 Oct;66(10):2263-5. doi: 10.1093/jac/dkr310. Epub 2011 Jul 26.
Linked bla(CTX-M-3)-bla(TEM-1b) genes, as found on the IncI1 plasmid pEK204 prevalent in Belfast, also occur on plasmids belonging to other rep types in Escherichia coli isolated from nursing-home residents in Belfast. We investigated the mechanisms for their joint dissemination among diverse plasmids.
Plasmid pEK204 was transferred by electroporation into E. coli DH5α harbouring derivative pBAD Myc-His vectors. Transposition experiments were then performed at 37°C. Transposition of bla genes onto the derivative pBAD Myc-His vector was confirmed by sequencing.
ISEcp1 mediated transposition of bla(CTX-M-3) alone from pEK204, as well as both bla(CTX-M-3) and bla(TEM-1b) jointly. The 5' and 3' termini of the transposed fragments were identical to or resembled the ISEcp1 IR(L) and IR(R), respectively, thereby replicating the environments previously found in various Belfast clinical plasmids harbouring bla(CTX-M-3).
Simultaneous dissemination of bla(CTX-M-3) and bla(TEM-1b) among plasmids in Belfast's nursing homes is facilitated by ISEcp1-mediated transposition of these bla genes from pEK204-like and other plasmids. Such transposition events are of public health concern, as they potentially allow wider dissemination of CTX-M-3 enzyme than would be possible through the spread of extended-spectrum β-lactamase-encoding plasmids and strains alone.
在贝尔法斯特流行的 IncI1 质粒 pEK204 上发现的 bla(CTX-M-3)-bla(TEM-1b) 基因也存在于其他 rep 类型的质粒中,这些质粒在贝尔法斯特的养老院居民中分离的大肠杆菌中。我们研究了它们在不同质粒中共同传播的机制。
通过电穿孔将质粒 pEK204 转移到携带衍生 pBAD Myc-His 载体的大肠杆菌 DH5α 中。然后在 37°C 下进行转座实验。通过测序确认 bla 基因转移到衍生的 pBAD Myc-His 载体上。
ISEcp1 介导 bla(CTX-M-3) 基因从 pEK204 单独转座,以及 bla(CTX-M-3) 和 bla(TEM-1b) 基因共同转座。转座片段的 5' 和 3' 末端与 ISEcp1 的 IR(L) 和 IR(R) 相同或相似,从而复制了先前在携带 bla(CTX-M-3) 的各种贝尔法斯特临床质粒中发现的环境。
ISEcp1 介导的这些 bla 基因从 pEK204 样和其他质粒中的转座,促进了贝尔法斯特养老院中质粒之间 bla(CTX-M-3) 和 bla(TEM-1b) 的同时传播。这种转座事件引起了公共卫生关注,因为它们可能会导致 CTX-M-3 酶的传播范围比通过扩展型β-内酰胺酶编码质粒和菌株的传播更广泛。