Transposition of ISEcp1 modulates blaCTX-M-55-mediated Shigella flexneri resistance to cefalothin.

The aim of this study was to uncover the mechanisms underlying Shigella flexneri resistance to cefalothin. In this study, a resistance-related S. flexneri isolate, S. flexneri YDC, was obtained from S. flexneri mel-1998023/zz pre-incubated with cefalothin at a dose of 0.5 × the minimum inhibitory concentration. The ISEcp1 coding element was identified upstream of bla(CTX-M-55) in S. flexneri YDC. To further determine the role of ISEcp1 in S. flexneri resistance, plasmids containing bla(CTX-M-55) recombinant with or without the ISEcp1 sequence were constructed and named as pCTX and pISECTX, respectively. It was shown that Escherichia coli DH5α(pISECTX) was resistant to all β-lactams tested. In contrast, E. coli DH5α(pCTX) was sensitive to all except β-lactams cefazolin and cefalothin. In addition, reverse transcription PCR showed that expression levels of bla(CTX-M-55) were higher in E. coli DH5α(pISECTX). The Clinical and Laboratory Standards Institute (CLSI) assay demonstrated that extended-spectrum β-lactamase was only positively detected in E. coli DH5α(pISECTX) but not in E. coli DH5α(pCTX). Taken together, these results suggest that the translocated ISEcp1 element upstream of bla(CTX-M-55) is required for overexpression of bla(CTX-M-55), leading to cephalosporin resistance.

Wang Y ，Song C ，Duan G ，Zhu J ，Yang H ，Xi Y ，Fan Q ... -  《-》

Occurrence, prevalence and genetic environment of CTX-M beta-lactamases in Enterobacteriaceae from Indian hospitals.

Ensor VM ，Shahid M ，Evans JT ，Hawkey PM ... -  《JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY》

ISEcp1 element in association with bla(CTX-M) genes of E. coli that produce extended-spectrum β-lactamase among the elderly in community settings.

This study analyzed the relationship between the ISEcp1 element and bla(CTX-M) genes of Escherichia coli isolates that produce extended-spectrum β-lactamase (ESBL) in community settings. Nineteen E. coli isolates that produced CTX-M-type β-lactamase were collected from four communities of elderly people in Shenyang, China. Polymerase chain reaction (PCR) amplification and direct sequencing were used to detect the insertion of the ISEcp1 element into the genetic environment of the bla(CTX-M) genes. The ISEcp1 element was associated with several bla(CTX-M) gene types, including CTX-M-14, CTX-M-24, CTX-M-22, and CTX-M-79. Sequence analysis revealed that all of the ISEcp1-like DNA sequences contained the putative promoter region that is involved in CTX-M genes transcription. ISEcp1 insertion sequences were observed 42-127bp upstream of the open reading frames (ORFs) that encode the CTX-M enzymes in all 15 strains. The CTX-M-79 β-lactamase-encoding gene was observed with a different ISEcp1 insertion site and variable sequences between the ISEcp1 and bla(CTX-M-79) gene. For one strain (T298), the ISEcp1 element was disrupted by IS10. This work confirmed that the ISEcp1 elements were closely linked to bla(CTX-M) genes in community isolates from Shenyang, China.

Tian SF ，Chu YZ ，Chen BY ，Nian H ，Shang H ... -  《-》

Different IncI1 plasmids from Escherichia coli carry ISEcp1-blaCTX-M-15 associated with different Tn2-derived elements.

The bla(CTX-M-15) gene, encoding the globally dominant CTX-M-15 extended-spectrum β-lactamase, has generally been found in a 2.971-kb ISEcp1-bla(CTX-M-15)-orf477Δ transposition unit, with ISEcp1 providing a promoter. In available IncF plasmid sequences from Escherichia coli, this transposition unit interrupts a truncated copy of transposon Tn2 that lies within larger multiresistance regions. In E. coli, bla(CTX-M-15) is also commonly associated with IncI1 plasmids and here three such plasmids from E. coli clinical isolates from western Sydney 2006-2007 have been sequenced. The plasmid backbones are organised similarly to those of other IncI1 plasmids, but have insertions and/or deletions and sequence differences. Each plasmid also has a different insertion carrying bla(CTX-M-15). pJIE113 (IncI1 sequence type ST31) is almost identical to plasmids isolated from the 2011 E. coli O104:H4 outbreak in Europe, where the typical bla(CTX-M-15) transposition unit interrupts a complete Tn2 inserted directly in the plasmid backbone. In the novel plasmid pJIE139 (ST88), ISEcp1-blaC(TX-M-15)-orf477Δ lies within a Tn2/3 hybrid transposon. Homologous recombination could explain movement of ISEcp1-bla(CTX-M-15)-orf477Δ between copies of Tn2 on IncF and IncI1 plasmids and generation of the Tn2/3 hybrid. pJIE174 (ST37) is almost identical to pESBL-12 from the Netherlands and in these plasmids bla(CTX-M-15) is flanked by two copies of IS26 that truncate the transposition unit within a larger region bounded by the ends of Tn2. bla(CTX-M-15) and the associated ISEcp1-derived promoter may be able to move from this structure by the actions of IS26, independently of both ISEcp1 and Tn2.

Zong Z ，Ginn AN ，Dobiasova H ，Iredell JR ，Partridge SR ... -  《-》

ISEcp1-mediated transposition of linked blaCTX-M-3 and blaTEM-1b from the IncI1 plasmid pEK204 found in clinical isolates of Escherichia coli from Belfast, UK.

Dhanji H ，Doumith M ，Hope R ，Livermore DM ，Woodford N ... -  《-》