Molecular characterization of multidrug resistant hospital isolates using the antimicrobial resistance determinant microarray

• Published in: PloS one Vol. 8; no. 7; p. e69507
• Main Authors: Leski, Tomasz A, Vora, Gary J, Barrows, Brian R, Pimentel, Guillermo, House, Brent L, Nicklasson, Matilda, Wasfy, Momtaz, Abdel-Maksoud, Mohamed, Taitt, Chris Rowe
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 25.07.2013; Public Library of Science (PLoS)
• Subjects: Acinetobacter baumannii; Acinetobacter baumannii - drug effects; Acinetobacter baumannii - enzymology; Acinetobacter baumannii - genetics; Acinetobacter baumannii - isolation & purification; Acinetobacter Infections - drug therapy; Acinetobacter Infections - epidemiology; Acinetobacter Infections - microbiology; Algorithms; Amines; Analysis; Anti-Bacterial Agents - pharmacology; Antibiotic resistance; Antibiotics; Antiinfectives and antibacterials; Antimicrobial agents; Antimicrobial resistance; Bacteria; beta-Lactamases - genetics; beta-Lactamases - metabolism; Biology; Deoxyribonucleic acid; Determinants; DNA; DNA microarrays; DNA sequencing; DNA, Bacterial - analysis; DNA, Bacterial - genetics; Drug resistance; Drug Resistance, Multiple, Bacterial - genetics; E coli; Egypt - epidemiology; Enterobacteriaceae; Enterococcus faecalis; Epidemiology; Escherichia coli; Escherichia coli - drug effects; Escherichia coli - enzymology; Escherichia coli - genetics; Escherichia coli - isolation & purification; Escherichia coli Infections - drug therapy; Escherichia coli Infections - epidemiology; Escherichia coli Infections - microbiology; Evolution; Genes; Genomes; Genomics; Humans; Klebsiella; Klebsiella Infections - drug therapy; Klebsiella Infections - epidemiology; Klebsiella Infections - microbiology; Klebsiella pneumoniae; Klebsiella pneumoniae - drug effects; Klebsiella pneumoniae - enzymology; Klebsiella pneumoniae - genetics; Klebsiella pneumoniae - isolation & purification; Laboratories; Medical research; Medicine; Microbial drug resistance; Multidrug resistance; Multidrug resistant organisms; Nucleotide sequence; Oligonucleotide Array Sequence Analysis - methods; Polymerase Chain Reaction; Quaternary; Sequence Analysis, DNA; Staphylococcus aureus; Staphylococcus epidermidis; Staphylococcus infections; Strains (organisms); Streptothricin; Urogenital system; Egypt; Washington DC; United States > US; Maryland
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0069507

Molecular methods that enable the detection of antimicrobial resistance determinants are critical surveillance tools that are necessary to aid in curbing the spread of antibiotic resistance. In this study, we describe the use of the Antimicrobial Resistance Determinant Microarray (ARDM) that targets 239 unique genes that confer resistance to 12 classes of antimicrobial compounds, quaternary amines and streptothricin for the determination of multidrug resistance (MDR) gene profiles. Fourteen reference MDR strains, which either were genome, sequenced or possessed well characterized drug resistance profiles were used to optimize detection algorithms and threshold criteria to ensure the microarray's effectiveness for unbiased characterization of antimicrobial resistance determinants in MDR strains. The subsequent testing of Acinetobacter baumannii, Escherichia coli and Klebsiella pneumoniae hospital isolates revealed the presence of several antibiotic resistance genes [e.g. belonging to TEM, SHV, OXA and CTX-M classes (and OXA and CTX-M subfamilies) of β-lactamases] and their assemblages which were confirmed by PCR and DNA sequence analysis. When combined with results from the reference strains, ~25% of the ARDM content was confirmed as effective for representing allelic content from both Gram-positive and -negative species. Taken together, the ARDM identified MDR assemblages containing six to 18 unique resistance genes in each strain tested, demonstrating its utility as a powerful tool for molecular epidemiological investigations of antimicrobial resistance in clinically relevant bacterial pathogens.