Development of a single-reaction multiplex PCR toxin typing assay for Staphylococcus aureus strains.
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.
Sharma
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NK
Division of Food Sciences, School of Biological Sciences, University of Nottingham, Sutton Bonington Campus, Loughborough LE12 5RD, United Kingdom.
Rees
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CE
Dodd
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CE
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*
D017931:DNA Primers; D004768:Enterotoxins