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Applied and Environmental Microbiology, April 2000, p. 1347-1353, Vol. 66, No. 4
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Development of a Single-Reaction Multiplex PCR Toxin Typing Assay for Staphylococcus aureus Strains

Naresh K. Sharma, Catherine E. D. Rees, and Christine E. R. Dodd*

Division of Food Sciences, School of Biological Sciences, University of Nottingham, Sutton Bonington Campus, Loughborough LE12 5RD, United Kingdom

Received 21 October 1999/Accepted 1 January 2000

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.

* Corresponding author. Mailing address: Division of Food Sciences, School of Biological Sciences, University of Nottingham, Sutton Bonington Campus, Loughborough LE12 5RD, United Kingdom. Phone: 44(0)1159516163. Fax: 44(0)1159516162. E-mail: christine.dodd{at}nottingham.ac.uk.

Applied and Environmental Microbiology, April 2000, p. 1347-1353, Vol. 66, No. 4
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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