title: Screening of large numbers of Streptococcus pneumoniae isolates for mutations associated with fluoroquinolone resistance using an oligonucleotide probe assay.
abstract: Mutations at a relatively small number of sites in parC, parE and gyrA account for most of the fluoroquinolone resistance in Streptococcus pneumoniae clinical isolates. A high throughput oligonucleotide probe assay was developed to screen for mutations in the quinolone-resistance determining region (QRDR) of parC (Ser79), gyrA (Ser81) and parE (Asp435) of Streptococcus pneumoniae. Eight oligonucleotide probes (17mers) were used in the presence of tetramethyl ammonium chloride so that the melting temperature was dependent on length and not on base composition. Using this assay it was possible to accurately detect QRDR mutations from several hundred S. pneumoniae clinical isolates that were grown on nylon membranes.
authors.affiliation: Johnson and Johnson Pharmaceutical Research and Development, LLC, 1000 Route 202, Raritan, NJ 08869, USA. tdavies@prdus.jnj.com
authors.forename: Todd A
authors.initials: TA
authors.lastname: Davies
authors.forename: Raul
authors.initials: R
authors.lastname: Goldschmidt
mesh_terms: D000890:Anti-Infective Agents / Q000494:pharmacology*; D001483:Base Sequence; D004252:DNA Mutational Analysis / Q000379:methods*; D024881:Drug Resistance, Bacterial / Q000235:genetics; D024841:Fluoroquinolones; D005798:Genes, Bacterial; D009154:Mutation; D009693:Nucleic Acid Hybridization / Q000379:methods; D015345:Oligonucleotide Probes; D013296:Streptococcus pneumoniae / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D000890:Anti-Infective Agents; D024841:Fluoroquinolones; D015345:Oligonucleotide Probes