title: Comparison of two DNA microarrays for detection of plasmid-mediated antimicrobial resistance and virulence factor genes in clinical isolates of Enterobacteriaceae and non-Enterobacteriaceae.
abstract: A DNA microarray was developed to detect plasmid-mediated antimicrobial resistance (AR) and virulence factor (VF) genes in clinical isolates of Enterobacteriaceae and non-Enterobacteriaceae. The array was validated with the following bacterial species: Escherichiacoli (n=17); Klebsiellapneumoniae (n=3); Enterobacter spp. (n=6); Acinetobacter genospecies 3 (n=1); Acinetobacterbaumannii (n=1); Pseudomonasaeruginosa (n=2); and Stenotrophomonasmaltophilia (n=2). The AR gene profiles of these isolates were identified by polymerase chain reaction (PCR). The DNA microarray consisted of 155 and 133 AR and VF gene probes, respectively. Results were compared with the commercially available Identibac AMR-ve Array Tube. Hybridisation results indicated that there was excellent correlation between PCR and array results for AR and VF genes. Genes conferring resistance to each antibiotic class were identified by the DNA array. Unusual resistance genes were also identified, such as bla(SHV-5) in a bla(OXA-23)-positive carbapenem-resistant A. baumannii. The phylogenetic group of each E. coli isolate was verified by the array. These data demonstrate that it is possible to screen simultaneously for all important classes of mobile AR and VF genes in Enterobacteriaceae and non-Enterobacteriaceae whilst also assigning a correct phylogenetic group to E. coli isolates. Therefore, it is feasible to test clinical Gram-negative bacteria for all known AR genes and to provide important information regarding pathogenicity simultaneously.
authors.affiliation: Department of Clinical Microbiology, Sir Patrick Dun Translational Research Laboratory, School of Medicine, University of Dublin, Trinity College, St James's Hospital Campus, Dublin 8, Ireland. fiona1walsh@gmail.com
authors.forename: Fiona
authors.initials: F
authors.lastname: Walsh
authors.forename: Niamh M
authors.initials: NM
authors.lastname: Cooke
authors.forename: Stephen G
authors.initials: SG
authors.lastname: Smith
authors.forename: Gary P
authors.initials: GP
authors.lastname: Moran
authors.forename: Fiona J
authors.initials: FJ
authors.lastname: Cooke
authors.forename: Alasdair
authors.initials: A
authors.lastname: Ivens
authors.forename: John
authors.initials: J
authors.lastname: Wain
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Rogers
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D001431:Bacteriological Techniques / Q000379:methods*; D024881:Drug Resistance, Bacterial; D005798:Genes, Bacterial; D006090:Gram-Negative Bacteria / Q000235:genetics* / Q000302:isolation & purification; D016905:Gram-Negative Bacterial Infections / Q000382:microbiology*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012680:Sensitivity and Specificity; D037521:Virulence Factors / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D037521:Virulence Factors