title: DNA microarray for detection of macrolide resistance genes.
abstract: A DNA microarray was developed to detect bacterial genes conferring resistance to macrolides and related antibiotics. A database containing 65 nonredundant genes selected from publicly available DNA sequences was constructed and used to design 100 oligonucleotide probes that could specifically detect and discriminate all 65 genes. Probes were spotted on a glass slide, and the array was reacted with DNA templates extracted from 20 reference strains of eight different bacterial species (Streptococcus pneumoniae, Streptococcus pyogenes, Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, Staphylococcus haemolyticus, Escherichia coli, and Bacteroides fragilis) known to harbor 29 different macrolide resistance genes. Hybridization results showed that probes reacted with, and only with, the expected DNA templates and allowed discovery of three unexpected genes, including msr(SA) in B. fragilis, an efflux gene that has not yet been described for gram-negative bacteria.
authors.affiliation: LAMMB, Università di Siena, Policlinico Le Scotte/V Lotto, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Cassone
authors.forename: Marco M
authors.initials: MM
authors.lastname: D'Andrea
authors.forename: Francesco
authors.initials: F
authors.lastname: Iannelli
authors.forename: Marco R
authors.initials: MR
authors.lastname: Oggioni
authors.forename: Gian Maria
authors.initials: GM
authors.lastname: Rossolini
authors.forename: Gianni
authors.initials: G
authors.lastname: Pozzi
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D015342:DNA Probes; D004269:DNA, Bacterial; D030541:Databases, Genetic; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D005798:Genes, Bacterial; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000235:genetics; D006094:Gram-Positive Bacteria / Q000187:drug effects / Q000235:genetics; D018942:Macrolides / Q000494:pharmacology*; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D013698:Templates, Genetic
chemical_list: D000900:Anti-Bacterial Agents; D015342:DNA Probes; D004269:DNA, Bacterial; D018942:Macrolides