Detection of Escherichia coli heat-stable enterotoxin genes in pig stool specimens by an immobilized, colorimetric, nested polymerase chain reaction.

Journal of clinical microbiology ·Vol. 29 ·No. 11 ·1991-11-00 ·Pages 2375-9

Hornes E, Wasteson Y, Olsvik O

Abstract

A combination of selective enrichment by using immunomagnetic separation of F4 (K88)-positive Escherichia coli and a nested colorimetric polymerase chain reaction (PCR) was used on crude clinical and spiked samples for determination of genes encoding heat-stable enterotoxins (STs) Ia (ST Ia) and Ib (ST Ib). The combination increased the sensitivity of the nested PCR compared with that of application onto crude samples. Dead cells were also enriched by use of this technology, giving results that are not available by traditional cultivation as enrichment before PCR. The second step in the PCR was modified to be able to differentiate between ST Ia and ST Ib genes. The colorimetric PCR was performed in a microtiter format, making it useful for automation in clinical laboratories and for the screening of large numbers of samples.

MeSH Terms

Animals Bacterial Toxins/genetics Base Sequence Colorimetry DNA Probes DNA, Bacterial/genetics Enterotoxins/genetics Escherichia coli/genetics,isolation & purification,pathogenicity Escherichia coli Proteins Evaluation Studies as Topic Feces/microbiology Genes, Bacterial Molecular Sequence Data Polymerase Chain Reaction/methods,statistics & numerical data Sensitivity and Specificity Swine/microbiology

Chemicals

Bacterial Toxins DNA Probes DNA, Bacterial Enterotoxins Escherichia coli Proteins heat stable toxin (E coli)

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Hornes E

Department of Microbiology and Immunology, Norwegian College of Veterinary Medicine, Oslo.

Wasteson Y

Olsvik O

References (22)

22 references, click to expand

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Article Info

Journal

Journal of clinical microbiology

Abbr.

J Clin Microbiol

ISSN

0095-1137

DOI

10.1128/jcm.29.11.2375-2379.1991

Published

1991-11-00

Pages

2375-9

Language

English

Region

United States

NLM ID

7505564

PMCID

PMC270341

Subset

External Links

PubMed source DOI: 10.1128/jcm.29.11.2375-2379.1991 PMC Full Text View journal details

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Detection of Escherichia coli heat-stable enterotoxin genes in pig stool specimens by an immobilized, colorimetric, nested polymerase chain reaction.

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