Linkage of acquired quinolone resistance (qnrS1).pdfVIP

Linkage of acquired quinolone resistance (qnrS1) and metallo-b-lactamase (blaVIM-1) genes in multiple species of Enterobacteriaceae from Bolzano, Italy Richard Aschbacher1, Michel Doumith2, David M. Livermore2, Clara Larcher1 and Neil Woodford2* 1Laboratorio Interaziendale di Microbiologia e Virologia, Via Amba Alagi 5, I-39100 Bolzano, Italy; 2Antibiotic Resistance Monitoring and Reference Laboratory, Health Protection Agency, Centre for Infections, 61 Colindale Avenue, London NW9 5EQ, UK Received 12 October 2007; returned 24 November 2007; revised 30 November 2007; accepted 2 December 2007 Objectives: Twenty-four of 209 oxyimino-cephalosporin- and/or aztreonam-resistant Enterobacteriaceae collected around Bolzano had reduced susceptibility or resistance to carbapenems and gave positive metallo-b-lactamase (MBL) tests. Their resistance mechanisms were investigated. Methods: Resistances were identified by Vitek 2 and MIC tests and isolates were genotyped by PFGE. Resistance genes were identified by PCR and sequencing, and plasmids were transferred by conju- gation and/or transformation. Plasmid-borne genes were identified by Southern blotting, and their genetic surroundings were investigated by PCR mapping. Results: The 24 isolates with positive EDTA/imipenem synergy tests had blaVIM-1 carried on 40–150 kb plasmids. Imipenem MICs ranged from 2 to 32 mg/L, while those of meropenem and ertapenem were lower. The isolates included a clonal cluster of 10 Klebsiella pneumoniae, two other K. pneumoniae isolates, and diverse isolates of Escherichia coli (seven), Klebsiella oxytoca (three) and Citrobacter freundii (two). Six MBL producers were aztreonam-susceptible; the 18 aztreonam-resistant isolates had co-resident extended-spectrum b-lactamases. blaVIM-1 occurred as the first cassette in class 1 inte- grons, with aacA4 as the second cassette. Quinolone resistance gene qnrS1 was detected in 21 of 24 (87.5%) blaVIM-1-positive isolates versus 14 of 185 (7.6%) blaVIM-negative