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 0.5x 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 DH5 alpha(pISECTX) was resistant to all beta-lactams tested. In contrast, E. coli DH5 alpha(pCTX) was sensitive to all except beta-lactams cefazolin and cefalothin. In addition, reverse transcription PCR showed that expression levels of bla(CTX-M-55) were higher in E. coli DH5 alpha(pISECTX). The Clinical and Laboratory Standards Institute (CLSI) assay demonstrated that extended-spectrum beta-lactamase was only positively detected in E. coli DH5 alpha(pISECTX) but not in E. coli DH5 alpha(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. (C) 2013 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.