title: Multiplex PCR for direct detection of Shiga toxigenic Escherichia coli strains producing the novel subtilase cytotoxin.
abstract: We have recently described a novel AB5 subtilase cytotoxin produced by certain Shiga toxigenic Escherichia coli (STEC) strains. This potentially lethal toxin may contribute to severe gastrointestinal and systemic disease in humans. In this study we have developed a trivalent PCR assay for the detection of the novel toxin A subunit gene subA, as well as stx1 and stx2. The three primer pairs used in the assay do not interfere with each other and generate amplification products of 556, 180, and 255 bp, respectively. The assay can be used for determining the toxin genotype of STEC isolates, as well as for direct detection of toxin genes in primary fecal culture extracts.
authors.affiliation: School of Molecular and Biomedical Science, University of Adelaide, Adelaide, S.A. 5005, Australia. adrienne.paton@adelaide.edu.au
authors.forename: Adrienne W
authors.initials: AW
authors.lastname: Paton
authors.forename: James C
authors.initials: JC
authors.lastname: Paton
mesh_terms: D003603:Cytotoxins / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D003967:Diarrhea / Q000175:diagnosis* / Q000382:microbiology; D004926:Escherichia coli / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004927:Escherichia coli Infections / Q000175:diagnosis / Q000382:microbiology; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005243:Feces / Q000382:microbiology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D021122:Protein Subunits / Q000235:genetics* / Q000378:metabolism; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D022622:Shiga Toxin 1 / Q000235:genetics / Q000378:metabolism; D022641:Shiga Toxin 2 / Q000235:genetics / Q000378:metabolism; D022603:Shiga Toxins / Q000235:genetics* / Q000378:metabolism
chemical_list: D003603:Cytotoxins; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D021122:Protein Subunits; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2; D022603:Shiga Toxins; D012697:Serine Endopeptidases