Phenotypic and genotypic characterization of Acinetobacter spp. panel strains: A cornerstone to facilitate antimicrobial development

Roshan D'Souza, Naina A. Pinto, Nguyen Le Phuong, Paul G. Higgins, Thao Nguyen Vu, Jung Hyun Byun, Young Lag Cho, Jong Rak Choi, Dongeun Yong

Research output: Contribution to journalArticle

Abstract

Acinetobacter spp. have emerged as significant pathogens causing nosocomial infections. Treatment of these pathogens has become a major challenge to clinicians worldwide, due to their increasing tendency to antibiotic resistance. To address this, much revenue and technology are currently being dedicated toward developing novel drugs and antibiotic combinations to combat antimicrobial resistance. To address this issue, we have constructed a panel of Acinetobacter spp. strains expressing different antimicrobial resistance determinants such as narrow spectrum β-lactamases, extended-spectrum β-lactamases, OXA-type-carbapenemase, metallo-beta-lactamase, and over-expressed AmpC β-lactamase. Bacterial strains exhibiting different resistance phenotypes were collected between 2008 and 2013 from Severance Hospital, Seoul. Antimicrobial susceptibility was determined according to the CLSI guidelines using agar dilution method. Selected strains were sequenced using Ion Torrent PGM system, annotated using RAST server and analyzed using Geneious pro 8.0. Genotypic determinants, such as acquired resistance genes, changes in the expression of efflux pumps, mutations, and porin alternations, contributing to the relevant expressed phenotype were characterized. Isolates expressing ESBL phenotype consisted of blaPER-1 gene, the overproduction of intrinsic AmpC beta-lactamase associated with ISAba1 insertion, and carbapenem resistance associated with production of carbapenem-hydrolyzing Ambler class D β-lactamases, such as OXA-23, OXA-66, OXA-120, OXA-500, and metallo-β-lactamase, SIM-1. We have analyzed the relative expression of Ade efflux systems, and determined the sequences of their regulators to correlate with phenotypic resistance. Quinolone resistance-determining regions were analyzed to understand fluoroquinolone-resistance. Virulence factors responsible for pathogenesis were also identified. Due to several mutations, acquisition of multiple resistance genes and transposon insertion, phenotypic resistance decision scheme for for evaluating the resistance proved inaccurate, which highlights the urgent need for modification to this scheme. This complete illustration of mechanism contributing to specific resistance phenotypes can be used as a target for novel drug development. It can also be used as a reference strain in the clinical laboratory and for the evaluation of antibiotic efficacy for specific resistance mechanisms.

Original languageEnglish
Article number559
JournalFrontiers in Microbiology
Volume10
Issue numberMAR
DOIs
Publication statusPublished - 2019 Jan 1

Fingerprint

Acinetobacter
Phenotype
Carbapenems
Anti-Bacterial Agents
Porins
Mutation
Insertional Mutagenesis
Quinolones
Fluoroquinolones
Virulence Factors
Drug Combinations
beta-Lactamases
Microbial Drug Resistance
Cross Infection
Genes
Agar
Asp(5)-oxytocin
Guidelines
Ions
Technology

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Microbiology (medical)

Cite this

D'Souza, Roshan ; Pinto, Naina A. ; Le Phuong, Nguyen ; Higgins, Paul G. ; Vu, Thao Nguyen ; Byun, Jung Hyun ; Cho, Young Lag ; Choi, Jong Rak ; Yong, Dongeun. / Phenotypic and genotypic characterization of Acinetobacter spp. panel strains : A cornerstone to facilitate antimicrobial development. In: Frontiers in Microbiology. 2019 ; Vol. 10, No. MAR.
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Phenotypic and genotypic characterization of Acinetobacter spp. panel strains : A cornerstone to facilitate antimicrobial development. / D'Souza, Roshan; Pinto, Naina A.; Le Phuong, Nguyen; Higgins, Paul G.; Vu, Thao Nguyen; Byun, Jung Hyun; Cho, Young Lag; Choi, Jong Rak; Yong, Dongeun.

In: Frontiers in Microbiology, Vol. 10, No. MAR, 559, 01.01.2019.

Research output: Contribution to journalArticle

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T2 - A cornerstone to facilitate antimicrobial development

AU - D'Souza, Roshan

AU - Pinto, Naina A.

AU - Le Phuong, Nguyen

AU - Higgins, Paul G.

AU - Vu, Thao Nguyen

AU - Byun, Jung Hyun

AU - Cho, Young Lag

AU - Choi, Jong Rak

AU - Yong, Dongeun

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