Dataset Information

NDM-1 carbapenemase in Acinetobacter baumannii sequence type 32 in Ecuador.

ABSTRACT: Objectives:To describe a clinical case of Acinetobacter baumannii sequence type (ST) 32 harbouring a New Delhi metallo-?-lactamase (NDM) in Ecuador. Methods:We used multilocus sequence typing (MLST) to confirm the bacterial species and the sequence type of an A. baumannii isolate. We used synergy with the imipenem-EDTA disc method and the carbapenem inactivation method (CIM) to determine carbapenemase production; the presence of a carbapenemase gene was confirmed by PCR amplification and amplicon sequencing. Results:Molecular characterization revealed the presence of A. baumannii ST32 harbouring the bla NDM-1 gene in Ecuador. The bla NDM-1 gene was isolated through PCR and amplified from a purified plasmid. Conclusions:To the best of our knowledge, this is the first report of A. baumannii ST32 harbouring the bla NDM-1 gene.

SUBMITTER: Villacis JE

PROVIDER: S-EPMC6438911 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature

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NDM-1 carbapenemase in <i>Acinetobacter baumannii</i> sequence type 32 in Ecuador.

Villacís J E JE Bovera M M Romero-Alvarez D D Cornejo F F Albán V V Trueba G G Dorn H F HF Reyes J A JA

New microbes and new infections 20190308

<h4>Objectives</h4>To describe a clinical case of <i>Acinetobacter baumannii</i> sequence type (ST) 32 harbouring a New Delhi metallo-β-lactamase (NDM) in Ecuador.<h4>Methods</h4>We used multilocus sequence typing (MLST) to confirm the bacterial species and the sequence type of an <i>A. baumannii</i> isolate. We used synergy with the imipenem-EDTA disc method and the carbapenem inactivation method (CIM) to determine carbapenemase production; the presence of a carbapenemase gene was confirmed by ...[more]

PMID: 30976430

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Project description:Purpose:A novel variant of OXA-23, named OXA-423, was identified in an Acinetobacter baumannii clinical isolate. The aim of this study was to analyse the resistance phenotype of OXA-423. Methods:The A. baumannii strain WY-0713 was isolated from an intensive care unit patient. PCR was used to detect the bla OXA-23-like genes. Amplifying, cloning and sequencing were performed for the complete bla OXA-23-like. The novel bla OXA-423 and its ancestor bla OXA-23 were cloned into the expression vector pET-28b(+), and transformed into E. coli Rosetta (DE3) for antibiotic susceptibility testing. SDS-PAGE, modified Hodge test and CarbaNP test were used for detecting the expression of OXA-423 and OXA-23. Results:PCR screening of A. baumannii WY-0713 was positive for bla OXA-23-like genes. Sequencing of the PCR product identi?ed a novel bla OXA-23-like, named bla OXA-423 which encoding OXA-423. OXA-423 differed from OXA-23 by a crucial amino acid substitution (Val128Ala). The V128A substitution was located at the conserved active-site motifs SAV of OXA-23. Antibiotic susceptibility testing performed using isogenic E. coli showed that the MICs of E. coli Rosetta (pET-OXA-423) for penicillins and carbapenems were lower (reduced MICs 4-fold to 16-fold) than that of E. coli Rosetta (pET-OXA-23). The MICs of cefotaxime, ceftazidime and aztreonam for both transformants remained the same as the acceptor strain. Moreover, OXA-423 was slightly inhibited by sulbactam, clavulanic acid and tazobactam. SDS-PAGE analysis showed that OXA-423 and OXA-23 were conspicuously expressed. Modified Hodge test and CarbaNP test were positive demonstrated both of them were functional. Conclusion:OXA-423, the first report of an amino acid substitution located at conserved active-site motifs of OXA-23, conferred lower MIC values of penicillins and carbapenems as compared with OXA-23, while without affecting the resistance pro?les of expanded-spectrum cephalosporins and aztreonam.

Dataset Information

NDM-1 carbapenemase in Acinetobacter baumannii sequence type 32 in Ecuador.

Publications

NDM-1 carbapenemase in <i>Acinetobacter baumannii</i> sequence type 32 in Ecuador.

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