Improved method for the routine identification of toxigenic Escherichia coli by DNA amplification of a conserved region of the heat-labile toxin A subunit

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dc.contributor.authorVictor T.
dc.contributor.authorDu Toit R.
dc.contributor.authorVan Zyl J.
dc.contributor.authorBester A.J.
dc.contributor.authorVan Helden P.D.
dc.date.accessioned2011-05-15T15:56:59Z
dc.date.available2011-05-15T15:56:59Z
dc.date.issued1991
dc.description.abstractThis report describes a DNA amplification procedure for routine identification of heat-labile-toxin-producing Escherichia coli. Two oligonucleotide primers were used in a polymerase chain reaction procedure to amplify a highly conserved region of the A subunit of the heat-labile enterotoxin gene. Amplifications were done directly on E. coli colonies from plates when Salmonella, Shigella, or parasite infections were excluded as agents of the severe diarrhea in the patients. The conditions for the polymerase chain reaction method were empirically determined, and the procedure is inexpensive, sensitive, and specific. Positive results can be obtained over a wide variation in bacterial numbers, with no inhibition of Thermus aquaticus DNA polymerase. Detection of the amplified product can be done by agarose gel electrophoresis, which is specific and sensitive enough for routine diagnosis of this pathogen in clinical isolates. If greater sensitivity and specificity are required, hybridization with 32P- or alkaline phosphatase-labeled oligonucleotide probes can be used. Our results suggest that heat-labile-toxin-producing E. coli is responsible for about 9% of nondiagnosed diarrhea cases in Tygerberg Hospital, Tygerberg, Republic of South Africa.
dc.description.versionArticle
dc.identifier.citationJournal of Clinical Microbiology
dc.identifier.citation29
dc.identifier.citation1
dc.identifier.issn951137
dc.identifier.urihttp://hdl.handle.net/10019.1/10146
dc.subjectescherichia coli enterotoxin
dc.subjectoligonucleotide
dc.subjectphosphorus 32
dc.subjectarticle
dc.subjectbacterium identification
dc.subjectescherichia coli
dc.subjectgene amplification
dc.subjectnonhuman
dc.subjectpriority journal
dc.subjectAlkaline Phosphatase
dc.subjectBacterial Toxins
dc.subjectBase Sequence
dc.subjectDiarrhea
dc.subjectDNA, Bacterial
dc.subjectElectrophoresis, Agar Gel
dc.subjectEnterotoxins
dc.subjectEscherichia coli
dc.subjectEscherichia coli Infections
dc.subjectGenes, Bacterial
dc.subjectHuman
dc.subjectInfant
dc.subjectMolecular Sequence Data
dc.subjectPhosphorus Radioisotopes
dc.subjectPolymerase Chain Reaction
dc.subjectReproducibility of Results
dc.subjectSensitivity and Specificity
dc.subjectBacteria (microorganisms)
dc.subjectEscherichia coli
dc.subjectSalmonella
dc.subjectShigella
dc.subjectThermus aquaticus
dc.titleImproved method for the routine identification of toxigenic Escherichia coli by DNA amplification of a conserved region of the heat-labile toxin A subunit
dc.typeArticle
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