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Applied and Environmental Microbiology, October 2003, p. 6327-6333, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.6327-6333.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Multiplex Real-Time PCR Method To Identify Shiga Toxin Genes stx1 and stx2 and Escherichia coli O157:H7/H- Serotype

Karen C. Jinneman,1,2* Ken J. Yoshitomi,1,2 and Stephen D. Weagant2

Seafood Products Research Center,1 Pacific Regional Laboratory Northwest, U.S. Food and Drug Administration, Bothell, Washington2

Received 21 March 2003/ Accepted 31 July 2003

A multiplex real-time PCR method to simultaneously detect the stx1 and stx2 genes of Shiga toxin-producing Escherichia coli and a unique conserved single-nucleotide polymorphism in the E. coli O157:H7/H- uidA gene has been developed. There is more than 98.6% sensitivity and 100% specificity for all three gene targets based on a panel of 138 isolates. The PCR efficiencies were >=1.89, and as few as 6 CFU/reaction could be detected.

* Corresponding author. Mailing address: Seafood Products Research Center, Pacific Regional Laboratory Northwest, U.S. Food and Drug Administration, 22201 23rd Dr. SE, Bothell, WA 98021. Phone: (425) 483-4871. Fax: (425) 483-4996. E-mail: Karen.Jinneman{at}fda.gov.

Applied and Environmental Microbiology, October 2003, p. 6327-6333, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.6327-6333.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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