Co-Carriage of blaKPC-2 and blaNDM-1 in Clinical Isolates of Pseudomonas aeruginosa Associated with Hospital Infections from India

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.

Detection of carbapenemase-producing Pseudomonas aeruginosa by phenotypic and genotypic methods in a tertiary care hospital of East India

“…bla VIM + bla NDM-1 was the most common combination (6/102, 5.88%), followed by bla VIM + bla GIM (3/102, 2.94%). The coproduction of bla VIM + bla NDM-1 had previously been described in 7.1% CRPA isolates by Ellapan et al [15] The coexistence of bla VIM + bla NDM-1 and bla KPC-2 and bla NDM-1 in CRPA isolates from India is also described previously by Paul et al [18,19] No carbapenem-resistant gene was detected in 38 (38/102, 37.25%) CRPA isolates in our study. Other carbapenem mechanisms such as efflux pump and porin loss could have contributed for carbapenemase resistance, which were not studied in the present study.…”

Occurrence of co-existing bla VIM-2 and bla NDM-1 in clinical isolates of Pseudomonas aeruginosa from India

“…Similarly, a report from India also has established its chromosomal location [ 19 ]. However, in our recent study we found presence of bla NDM-1 on plasmid DNA indicating a possible shift from one to another genetic location [ 20 ]. Future experiments will show which direction (chromosome to plasmid or vice versa) of the transfer took place in the organism.…”

Carriage of blaNDM-1 in Pseudomonas aeruginosa through multiple Inc type plasmids in a tertiary referral hospital of northeast India

“…After overnight incubation, plasmids were extracted by QIAprep Spin Miniprep Kit (Qiagen, Germany). Plasmids of bla NDM-1 were subjected to transformation by heat shock method10 using Escherichia coli JM107 as recipient. Transformants were selected on LB agar with 0.25 μg/ml of imipenem, which were then confirmed both by phenotypic as well as by PCR analysis.…”

“…A total of 18 different replicon types such as FIA, FIB, FIC, HI1, HI2, I1/Iγ, L/M, N, P, W, T, A/C, K, B/O, X, Y, F and FIIA were targeted as described previously11. The antibiotic susceptibility was done by Kirby-Bauer disc-diffusion method10 against antibiotics viz . piperacillin-tazobactam (100/10 μg), co-trimoxazole (25 μg), amikacin (30 μg), gentamicin (10 μg), ciprofloxacin (5 μg), polymixin B (300 units), netilmicin (30 μg), carbenicillin (100 μg) and faropenem (5 μg) (Hi-Media, Mumbai, India).…”