BackgroundMolecular analysis of carbapenem-resistant genes in Acinetobacter baumannii, an emerging pathogen, is less commonly reported from Nepal. In this study we determined the antibiotic susceptibility profile and genetic mechanism of carbapenem resistance in clinical isolates of A. baumannii. Methods A. baumannii were isolated from various clinical specimens and identified based on Gram staining, biochemical tests, and PCR amplification of organism specific 16S rRNA and bla OXA-51 genes. The antibiotic susceptibility testing was performed using disc diffusion and E-test method. Multiplex PCR assays were used to detect the following β-lactamase genes: four class D carbapenem hydrolyzing oxacillinases (bla OXA-51, bla OXA-23, bla OXA-24 and bla OXA-58). Uniplex PCRs were used to detect three class B metallo-β-lactamases genes (bla IMP, bla VIM and bla NDM-1), class C cephalosporin resistance genes (bla ADC), aminoglycoside resistance gene (aphA6), and ISAba1 of all isolates. Insertion sequence ISAba125 among NDM-1 positive strains was detected. Clonal relatedness of all isolates were analyzed using repetitive sequence-based PCR (rep-PCR).ResultsOf total 44 analyzed isolates, 97.7% (n = 43) were carbapenem-resistant A. baumannii (CR-AB) and 97.7% (n = 43) were multidrug resistant A. baumannii (MDR-AB). One isolate was detected to be extremely drug resistant A. baumannii (XDR-AB). All the isolates were fully susceptible to colistin (MICs < 2 μg/ml). The bla OXA-23 gene was detected in all isolates, while bla NDM-1 was detected in 6 isolates (13.6%). Insertion sequence, ISAba1 was detected in all of bla OXA-23 positive isolates. ISAba125 was detected in all bla NDM-1 positive strains. The bla ADC and aphA6 genes were detected in 90.1 and 40.1%, respectively. The rep-PCR of all isolates represented 7 different genotypes.ConclusionWe found high prevalence of CR-AB and MDR-AB with bla OXA-23 gene in a tertiary care hospital in Nepal. Systemic network surveillance should be established for monitoring and controlling the spread of these resistant strains.Electronic supplementary materialThe online version of this article (doi:10.1186/s13756-017-0180-5) contains supplementary material, which is available to authorized users.

ObjectivesThis study was carried out to determine the prevalence of metallo-β-lactamases (MBLs) producing Pseudomonas aeruginosa in imipenem-nonsusceptible isolates and to detect MBL-encoding genes among MBLs-positive isolates.ResultsMetallo-β-lactamases production was detected in 68.6% isolates of P. aeruginosa with reduced susceptibility to imipenem. The bla VIM-2 gene was detected in 75% isolates and bla IMP-1 was detected in 25% isolates. All MBLs-positive isolates were multidrug resistant with a high level of resistance to imipenem (MIC 16 to ≥ 32 µg/ml), meropenem (MIC 16 to ≥ 32 µg/ml), and ceftazidime (MIC 64 to ≥ 512 µg/ml). All MBL-positive isolates were susceptible (MIC ≤ 2 µg/ml) to colistin. We found high prevalence of MBL-producing P. aeruginosa. To our knowledge this is the first report of detection of bla VIM-2 and bla IMP-1 in P. aeruginosa from Nepal. This indicates the need for awareness to prevent the spreading of these resistant isolates in hospital setting.

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