Detection of Shiga-like toxin (stx1 and stx2), intimin (eaeA), and enterohemorrhagic Escherichia coli (EHEC) hemolysin (EHEC hlyA) genes in animal feces by multiplex PCR

A multiplex PCR was developed for the rapid detection of genes encoding Shiga toxins 1 and 2 (stx1 and stx2), intimin (eaeA), and enterohemolysin A (hlyA) in 444 fecal samples derived from healthy and clinically affected cattle, sheep, pigs, and goats. The method involved non-solvent-based extractio...

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Bibliographic Details
Published in:Applied and Environmental Microbiology 1999-02, Vol.65 (2), p.868-872
Main Authors: Fagan, P.K, Hornitzky, M.A, Bettelheim, K.A, Djordjevic, S.P
Format: Article
Language:English
Subjects:
Adhesins, Bacterial
Animals
Animals, Domestic - microbiology
Bacteria
Bacterial Outer Membrane Proteins - genetics
Bacterial Proteins - genetics
bacterial toxins
Bacterial Toxins - genetics
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biological and medical sciences
Carrier Proteins
Cattle
DNA, Bacterial - analysis
Escherichia coli - genetics
Escherichia coli - pathogenicity
Escherichia coli Infections - microbiology
Escherichia coli Infections - veterinary
Escherichia coli Proteins
Feces
Feces - microbiology
Fundamental and applied biological sciences. Psychology
Genes
Hemolysin Proteins - genetics
Methods
Microbiology
polymerase chain reaction
Polymerase Chain Reaction - methods
Shiga Toxins
Virulence
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Summary:A multiplex PCR was developed for the rapid detection of genes encoding Shiga toxins 1 and 2 (stx1 and stx2), intimin (eaeA), and enterohemolysin A (hlyA) in 444 fecal samples derived from healthy and clinically affected cattle, sheep, pigs, and goats. The method involved non-solvent-based extraction of nucleic acid from an aliquot of an overnight culture of feces in EC (modified) broth. The detection limit of the assay for both fecal samples and pure cultures was between 18 and 37 genome equivalents. stx1 and hlyA were the most commonly encountered virulence factors.
ISSN:0099-2240
1098-5336
DOI:10.1128/aem.65.2.868-872.1999