title: Development of a single-reaction multiplex PCR toxin typing assay for Staphylococcus aureus strains.
abstract: We describe here the development of a single-reaction multiplex PCR assay for the enterotoxin genes from Staphylococcus aureus that utilizes a universal toxin gene primer in combination with toxin-specific primers to amplify characteristic toxin gene products. In combination with a new DNA purification method, the assay can detect enterotoxin genes A to E from a pure culture within 3 to 4 h. The test was used to characterize a diverse set of environmental S. aureus isolates, and a 99% correlation with toxin typing using standard immunological tests was found. The design of the assay allows it to be extended to include both newly characterized and as-yet-unknown toxin genes.
authors.affiliation: Division of Food Sciences, School of Biological Sciences, University of Nottingham, Sutton Bonington Campus, Loughborough LE12 5RD, United Kingdom.
authors.forename: N K
authors.initials: NK
authors.lastname: Sharma
authors.forename: C E
authors.initials: CE
authors.lastname: Rees
authors.forename: C E
authors.initials: CE
authors.lastname: Dodd
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D002417:Cattle; D017931:DNA Primers; D004768:Enterotoxins / Q000145:classification* / Q000235:genetics*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000379:methods*; D015183:Restriction Mapping; D012680:Sensitivity and Specificity; D013203:Staphylococcal Infections / Q000382:microbiology; D013211:Staphylococcus aureus / Q000145:classification / Q000235:genetics / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D004768:Enterotoxins