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. 2003 Oct;69(10):6327-33.
doi: 10.1128/AEM.69.10.6327-6333.2003.

Multiplex real-time PCR method to identify Shiga toxin genes stx1 and stx2 and Escherichia coli O157:H7/H- serotype

Karen C Jinneman  1 , Ken J Yoshitomi, Stephen D Weagant
Affiliations

Multiplex real-time PCR method to identify Shiga toxin genes stx1 and stx2 and Escherichia coli O157:H7/H- serotype

Karen C Jinneman et al. Appl Environ Microbiol. 2003 Oct.

Abstract

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.

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Figures

FIG. 1.
FIG. 1.
Standard curve for the multiplex real-time PCR analysis for the stx1 and stx2 genes and the E. coli O157:H7/H− uidA SNP at position 93 tested with E. coli O157:H7 isolate EDL 933. The standard deviations are based on three PCR amplifications.
FIG. 2.
FIG. 2.
Average Ct values for amplifications of stx1, stx2, and the E. coli O157:H7/H− uidA SNP at position 93 for reactions run individually and in multiplex format for E. coli O157:H7 strain EDL 933, based on 14 replicates.
See this image and copyright information in PMC

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