Dissemination of class I integron carrying VIM-2 carbapenemase gene in Pseudomonas

Transcription

1 AAC Accepts, published online ahead of print on 4 March 2013 Antimicrob. Agents Chemother. doi: /aac Copyright 2013, American Society for Microbiology. All Rights Reserved Dissemination of class I integron carrying VIM-2 carbapenemase gene in Pseudomonas aeruginosa clinical isolates from intensive care unit of University Hospital of Annaba, Algeria 4 5 Meriem Touati 1,2, Seydina M. Diene 1, Mazouz Dekhil 3, Abdelghani Djahoudi 4, Abdelkarim Racherache 5, Jean-Marc Rolain 1 * Unité de recherche sur les maladies infectieuses et tropicales émergentes, URMITE CNRS- IRD, UMR 6236, Méditerranée infection, Faculté de Médecine et de Pharmacie, Aix-Marseille- Université, Marseille, France. 2 Département de Biochimie, Université BADJI MOKHTAR. Annaba 23000, Algérie. 3 Laboratoire central de Microbiologie, CHU de Annaba-Algérie 4 Laboratoire de Microbiologie, Faculté de Médecine, université Badji Mokhtar, Annaba- Algérie. 5 Service de Réanimation, CHU d Annaba, Algérie These authors contribute equally to the work * Corresponding author Phone: (33) Fax: (33) jean-marc.rolain@univ-amu.fr Keywords: multidrug resistance; carbapenemase; Pseudomonas aeruginosa; Algeria, bla VIM-2 gene. 20 Text Word count =

2 22 Sir, Over the last decade, the increase of carbapenem resistance in Pseudomonas aeruginosa has mostly been due to impermeability due to OprD loss and MBL production, including IMP, VIM, SPM, GIM, SIM, AIM-1, FIM-1, and NDM (1,5,12,15); also, active efflux and serine carbapenemases may contribute to carbapenem resistance in P. aeruginosa in some cases. VIM-1 was reported in the Mediterranean area in 1997 in a clinical isolate of P. aeruginosa in Verona, Italy (10) and in a clinical isolate of P. mosselii isolated in 1994 in Genoa, Italy (4), and a VIM-2 variant appeared in France in Marseille, France in 1996 (14). VIM-2 has now spread as the predominant MBL variant among P. aeruginosa in all European Mediterranean countries (1), but in north Africa, the presence of VIM-2 has only been reported recently in Tunisia (5,9,11). Here we report the first molecular characterization of VIM-2-producing P. aeruginosa clinical isolates from Algeria that harbored a novel class I integron that also contained two gene cassettes encoding resistance to aminoglycosides (aadb and aaca4). A total of 17 non-replicate imipenem-resistant P. aeruginosa clinical isolates (imipenem MIC > 8 µg/ml as determined by Etest) recovered from December 2010 to September 2011 in a surgical intensive care unit at the University Hospital of Annaba, Algeria, were screened by PCR for the presence of MBL encoding genes using primers previously described (3,6,18). Antibiotic susceptibility testing was performed using disk diffusion according to the guidelines of the Antibiogram Committee of the French Society for Microbiology (CA-SFM) ( breakpoints and results are summarized in Table 1. Imipenem-EDTA synergy test (19) was positive for 14 out of 17 strains that were PCR-positive for VIM-2 carbapenemase using universal primers; VIM-all-F : 5 - TGGTCTACATGACCGCGTCT-3 and VIM-all-R: 3 - CGACTGAGCGATTTGTGTG-5 with expected PCR size of 766-bp. The characterization of class I integrons as reported (6) showed a novel class I integron that 2

3 harbored the VIM-2 gene associated with two gene cassettes encoding aminoglycosides resistance (aadb and aaca4). However, the molecular investigation of the remaining three strains using primers previously described (12) revealed chromosomic mutations that created premature stop codons in the oprd encoding gene. Complete MLST (2) revealed that the 17 strains belong to 4 different ST types including ST162 (one isolate), ST654 (one isolate), ST1175 (one isolate), and a new Sequence type, ST1420 (14 isolates), recently submitted in PubMLST database ( from China (January, 2013). To the best of our knowledge, in North Africa, VIM-2 MBL-producing P. aeruginosa have only been reported in Tunisia (7,9,11). VIM-2-producing P. aeruginosa have been reported in other countries in Africa including one report in a tourist from Hungary who was hospitalized in Egypt (17), one single case of a patient who was transferred to Norway after being hospitalized in Ghana (16), 57 clinical isolates recovered in 2006 and 2007 in Kenya (13) and a report of 15 clinical isolates in South Africa (8). Our results demonstrate that national surveillance should be urgently implemented in Algeria to monitor and control the emergence and spread of carbapenemase-encoding genes. Data accession The sequence full of this class 1 integron has been deposited on GenBank database under accession number JX Acknowledgments The authors thank Linda Hadjadj and Maxime Pit for technical assistance. We thank American Journal Experts for English corrections. Conflict of interest: None to declare. 68 All authors have read and approved the manuscript. 3

4 69 Funding source: This work was partly funded by CNRS. 4

5 70 Table 1. Phenotypic and genotypic features of the 17 P. aeruginosa clinical isolates imipenem-resistant Date Patient IMP Sequence Strain Sex Source TIC TCC TZP CAZ GEN CIP FOS COL VIM-2 of isolation age (year) MIC (µg/ml) type (ST) 1 15/12/ M Bronchial aspirate 12 R R R R R S S S + ST /12/ F Blood 32 R R R R R S S S + ST /12/ months M Bronchial aspirate 12 R R R R R S S S + ST /02/ months M Bronchial aspirate 8 R R R R R R R S + ST /02/ M Bronchial aspirate 16 R R R R R R R S + ST /03/ days M Bronchial aspirate 32 R R R R S S R S + ST /03/ M Urinary catheter 12 R R R R R R S S + ST /03/ M Urinary catheter 32 R R R R S S R S + ST1420 Downloaded from /04/ M Urine 12 R R R R S S R S + ST /05/ M Pus 16 R R R R R S R S + ST /06/ M Urine 16 R R R R S R S S + ST a 22/06/ F Bronchial aspirate 32 R R R R R S S S - ST b 22/07/ M Bronchial aspirate 32 R R R R S S R S - ST /08/ M Wound 16 R R R R R S S S + ST on April 9, 2018 by guest

6 /08/ M Wound 12 R R R R S S R S + ST c 10/09/ M Urine 16 R R R R R R R S - ST /08/ M Wound 12 R R R R R S S S + ST1420 CAZ, ceftazidime; CIP, ciprofloxacin; COL, colistin; FOS, fosfomycin; GEN, gentamycin; TCC, ticarcillin/clavulanic acid; TIC, ticarcillin; TZP, piperacillin/tazobactam M, Male; F, Female; R, Resistant; S, Susceptible; MIC, Minimum Inhibitory Concentration determined by Etest; R, resistant; S, Susceptible a 529 CGA to TGA mutation leading to stop codon in oprd gene; b 193 TGGA to TG - A deletion leading to stop codon in oprd gene; c 757 TCG to TAG mutation leading to stop codon in oprd gene. Downloaded from 6 on April 9, 2018 by guest

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