Development of a DNA microarray for enterococcal species, virulence, and antibiotic resistance gene determinations among isolates from poultry

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DOIResolve DOI: http://doi.org/10.1128/AEM.00263-11
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TypeArticle
Journal titleApplied and Environmental Microbiology
Volume77
Issue8
Pages26252633; # of pages: 9
Subjectaccuracy assessment; Acids; analysis; Animal; Animals; antibiotic resistance; Antibiotic resistance genes; antibiotics; Antimicrobial resistance; array; Bacteria; bacterium; bio; Biotechnology; Broiler chickens; Canada; Chickens; crossector; discriminant analysis; Dna; DNA micro-array; DNA microarray; Dogs; Enterococcus faecium; env; Enzymes; Escherichia coli; Evolution; Fermentation; Gene Expression; Genes; Genomics; genotype; Human; Humans; Hybridization; identification method; immunology; methods; Microarray; Microarrays; microbiology; Microorganisms; monitoring; Mycoplasma; Nucleic Acids; Operon; pathogenicity; PCR; Phenotype; plastic; poultry; Probes; Protein; Scopus; Simultaneous identification; Species identification; Species Specificity; taxonomy; Teicoplanin; Virulence; Virulence factors; Virulence gene; Virulence potential
AbstractA DNA microarray (Enteroarray) was designed with probes targeting four species-specific taxonomic identifiers to discriminate among 18 different enterococcal species, while other probes were designed to identify 18 virulence factors and 174 antibiotic resistance genes. In total, 262 genes were utilized for rapid species identification of enterococcal isolates, while characterizing their virulence potential through the simultaneous identification of endogenous antibiotic resistance and virulence genes. Enterococcal isolates from broiler chicken farms were initially identified by using the API 20 Strep system, and the results were compared to those obtained with the taxonomic genes atpA, recA, pheS, and ddl represented on our microarray. Among the 171 isolates studied, five different enterococcal species were identified by using the API 20 Strep system: Enterococcus faecium, E. faecalis, E. durans, E. gallinarum, and E. avium. The Enteroarray detected the same species as API 20 Strep, as well as two more: E. casseliflavus and E. hirae. Species comparisons resulted in 15% (27 isolates) disagreement between the two methods among the five API 20 Strep identifiable species and 24% (42 isolates) disagreement when considering the seven Enteroarray identified species. The species specificity of key antibiotic and virulence genes identified by the Enteroarray were consistent with the literature adding further robustness to the redundant taxonomic probe data. Sequencing of the cpn60 gene further confirmed the complete accuracy of the microarray results. The new Enteroarray should prove to be a useful tool to accurately genotype strains of enterococci and assess their virulence potential. © 2011, American Society for Microbiology.
Publication date
LanguageEnglish
AffiliationNRC Biotechnology Research Institute; National Research Council Canada
Peer reviewedYes
NRC number52784
NPARC number18335889
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Record identifier062be0e3-fb81-4ee3-976a-9b8e16d26864
Record created2011-07-29
Record modified2017-03-23
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