Abstract
Global spread of KPC poses to be a serious threat complicating treatment options in hospital settings. The present study investigates the genetic environment of bla KPC-2 among clinical isolates of Pseudomonas aeruginosa from a tertiary referral hospital of India. The study isolates were collected from different wards and clinics of Silchar Medical College and Hospital, India, from 2012–2013. The presence of bla KPC was confirmed by genotypic characterization followed by sequencing. Cloning of the bla KPC-2 gene was performed and the genetic environment of this gene was characterized as well. Transferability of the resistance gene was determined by transformation assay and Southern hybridization. Additionally, restriction mapping was also carried out. Two isolates of P. aeruginosa were found to harbor bla KPC-2, were resistant towards aminoglycosides, quinolone and β-lactam-β-lactamase inhibitor combination. In both the isolates, the resistance determinant was associated with class 1 integron and horizontally transferable. Both the isolates were co-harboring bla NDM-1. The first detection of this integron mediated bla KPC-2 coexisting with bla NDM-1 in P. aeruginosa from India is worrisome, and further investigation is required to track the gene cassette mediated bla KPC-2 in terms of infection control and to prevent the spread of this gene in hospitals as well as in the community.
Highlights
Klebsiella pneumoniae carbapenemase (KPC) is a major contributor for carbapenem resistance across the globe within the members of Enterobacteriaceae [1] and in recent past they are been reported in Pseudomonas spp. from Brazil and China [2, 3]
Restriction analysis and sequencing revealed KPC-2 was present in two different locations within the host
59 base element PCR confirmed blaKPC-2 was flanked by attC site. blaKPC-2 was flanked by dfr16 and aadA2 in the upstream region while qac was located in the downstream area within the cassette
Summary
Klebsiella pneumoniae carbapenemase (KPC) is a major contributor for carbapenem resistance across the globe within the members of Enterobacteriaceae [1] and in recent past they are been reported in Pseudomonas spp. from Brazil and China [2, 3]. In Indian subcontinent, this gene has so far been reported only in Enterobacteriaceae family [4]. Co-Carriage of blaKPC-2 and blaNDM-1 in Clinical Isolates witnessed with other carbapenemase genes as well [4]. No study from this country has attempted to investigate genetic background of this gene. We have reported coexistence of blaKPC-2 and blaNDM-1with different genetic context in P. aeruginosa in a tertiary referral hospital of India
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