Evaluation of genetic determinants involved in β-lactam- and multiresistance in a surgical ICU

Antibiotic resistance is a major and well-known problem in intensive care units (ICUs) world-wide and previously susceptible isolates become resistant through the acquisition of resistance determinants from other bacteria or the development of mutations, as is the case in β-lactam resistance. We evaluated the presence of resistance determinants involved in β-lactam resistance and multi-resistance in order to establish the contribution of horizontal gene transfer to the spread of resistance in a surgical ICU during an antibiotic rotation study. Pseudomonas aeruginosa and Enterobacteriaceae isolates were selected and iso-electric focusing (IEF), DNA-typing methods such as specific β-lactamase and specific integron PCRs were performed to determine the presence of β-lactamases. The PCRs specific for IMP-1, OXA-1, and VIM-type β-lactamases performed on the selected P. aeruginosa and Enterobacteriaceae isolates with MICs for cephalosporins >1mg/l did not demonstrate any of these β-lactamases. IEF for 14 pseudomonads, representing 7 genotypes from 9 patients, showed a β-lactamase with a pI larger than 8.5 in 13 of the isolates. The integrase PCR was positive for only five isolates from three patients and conserved segment PCR showed integrons of variable sizes (700, 900, 1400 and 1500bp). Each patient had its own integron types. It can be concluded that integrons and associated resistance determinants played only a minor role in the surgical ICU and β-lactam resistance among P. aeruginosa isolates was most likely due to the derepression of its AmpC gene.