Multiplex real-time PCR method to identify Shiga toxin genes stx1 and stx2 and Escherichia coli O157:H7/H- serotype.
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.
Jinneman
Karen C
KC
Seafood Products Research Center, Pacific Regional Laboratory Northwest, U.S. Food and Drug Administration, Bothell, Washington 98021, USA. Karen.Jinneman@fda.gov
Yoshitomi
Ken J
KJ
Weagant
Stephen D
SD
D015373:Bacterial Typing Techniques; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000032:analysis; D004927:Escherichia coli Infections / Q000382:microbiology; D019453:Escherichia coli O157 / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D005966:Glucuronidase / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D012680:Sensitivity and Specificity; D012703:Serotyping; D022622:Shiga Toxin 1 / Q000235:genetics*; D022641:Shiga Toxin 2 / Q000235:genetics*
D004269:DNA, Bacterial; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2; D005966:Glucuronidase