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PMID: 3299267 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Enzyme-linked synthetic oligonucleotide probes: non-radioactive detection of enterotoxigenic Escherichia coli in faecal specimens.

Nucleic acids research ·Vol. 15 ·No. 13 ·1987-07-10 ·Pages 5275-87

Li P, Medon PP, Skingle DC, Lanser JA, Symons RH

Abstract

Synthetic oligonucleotides, complementary to unique sequences in the heat stable enterotoxin gene of Escherichia coli specific for humans, were prepared with a 30-atom spacer arm and a 3' terminal sulfhydryl group which was coupled to bromoacetyl-derivatized alkaline phosphatase. The resulting direct enzyme-linked oligonucleotide probes, containing one enzyme molecule per oligonucleotide, successfully diagnosed enterotoxigenic Escherichia coli in clinical specimens by using a modified colony hybridization method and a colorimetric assay. The procedure is rapid, simple and reliable with a sensitivity equivalent to that using 5'-terminally labelled [32p]-oligonucleotide probes. The results indicate that the enzyme-labelled oligonucleotide probes should be applicable to the routine diagnosis of enterotoxigenic Escherichia coli and possess the potential for the detection of other microbial pathogens.

MeSH Terms
Alkaline Phosphatase Australia Bacterial Toxins/analysis,genetics Diarrhea/microbiology Enterotoxins/analysis,genetics Escherichia coli/genetics,isolation & purification Escherichia coli Proteins Ethnicity Feces/microbiology Humans Native Hawaiian or Other Pacific Islander Nucleic Acid Hybridization Oligodeoxyribonucleotides/chemical synthesis
Chemicals
Bacterial Toxins Enterotoxins Escherichia coli Proteins Oligodeoxyribonucleotides heat stable toxin (E coli) Alkaline Phosphatase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Li P
Medon P P
Skingle D C
Lanser J A
Symons R H
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27 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/15.13.5275
Published
1987-07-10
Pages
5275-87
Language
English
Region
England
NLM ID
0411011
PMCID
PMC305961
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/15.13.5275 PMC Full Text View journal details
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