Co-existence of blaOXA-23 and blaNDM-1 genes of Acinetobacter baumannii isolated from Nepal: antimicrobial resistance and clinical significance

Prabhu Raj Joshi,Sutthirat Sitthisak,Mahesh Acharya,Trishna Kakshapati,Udomluk Leungtongkam,Rapee Thummeepak

doi:10.1186/s13756-017-0180-5

Abstract

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.MethodsA. baumannii were isolated from various clinical specimens and identified based on Gram staining, biochemical tests, and PCR amplification of organism specific 16S rRNA and blaOXA-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 (blaOXA-51, blaOXA-23, blaOXA-24 and blaOXA-58). Uniplex PCRs were used to detect three class B metallo-β-lactamases genes (blaIMP, blaVIM and blaNDM-1), class C cephalosporin resistance genes (blaADC), 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 blaOXA-23 gene was detected in all isolates, while blaNDM-1 was detected in 6 isolates (13.6%). Insertion sequence, ISAba1 was detected in all of blaOXA-23 positive isolates. ISAba125 was detected in all blaNDM-1 positive strains. The blaADC 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 blaOXA-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.

Highlights

Molecular analysis of carbapenem-resistant genes in Acinetobacter baumannii, an emerging pathogen, is less commonly reported from Nepal
The zones of inhibition were determined whether the microorganism was susceptible, intermediately resistant, or resistant to each antibiotic according to Clinical and Laboratory Standards Institute (CLSI) guidelines
Antibiotic susceptibility Of the 44 isolates, resistance was found against ciprofloxacin (n = 43, 97.7%), cefotaxime (n = 43, 97.7%), ceftazidime (n = 42, 95.4%), ceftriaxone (n = 41, 93.2%), cefepime (n = 39, 88.6%), amikacin (n = 19, 43.2%), gentamicin (n = 23, 52.3%), trimethoprim/sulfamethoxazole (n = 41, 93.2%), tetracycline (n = 21, 47.7%) and piperacillin/tazobactam (n = 43, 97.7%)

Summary

Introduction

Molecular 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. Acinetobacter baumannii, an emerging pathogen of healthcare centers, shows intrinsic as well as acquired drug-resistance mechanisms [1]. Acinetobacter spp. develop carbapenem resistance by production of OXA-type carbapenemase and metallo-β-lactamases (MBLs) [6, 7]; blaOXA23-like, blaOXA-40-like, blaOXA-58-like and blaOXA-51-like carbapenemases are broadly reported, where blaOXA-51like β-lactamases, intrinsic to A. baumannii, is used for species identification [8,9,10]. A. baumannii harboring plasmid encoded New Delhi metallo-β-lactamase-1 (NDM-1), a novel carbapenemase gene, is reported from many countries [11, 12]. Detection of class C β-lactamase genes (blaADC) which mediated cephalosporin resistances and aminoglycoside resistant genes (aphA6) has increased in recent years in A. baumannii clinical isolates [13, 14]

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