Document Detail

Multiplex PCR-DNA probe assay for the detection of pathogenic Escherichia coli.
MedLine Citation:
PMID:  15567229     Owner:  NLM     Status:  MEDLINE    
A multiplex PCR-DNA probing assay was developed to detect four major Escherichia coli virotypes. Six highly specific polymerase chain reaction (PCR) primer sets and DIG-labeled chemiluminescent probes were designed to target the Shiga-like toxin I and II genes (stxI and stxII) of verotoxigenic E. coli (VTEC), heat-stable and heat-labile toxin genes of enterotoxigenic E. coli (ETEC), adherence factor (EAF) of enteropathogenic E. coli (EPEC) and a fragment of the invasiveness plasmid (IAL) of enteroinvasive E. coli (EIEC). The primer pairs generate products of 350, 262, 170, 322, 293 and 390 bp in length, respectively. The multiplex primers and probes were tested for specificity against 31 pathogenic E. coli strains, nine nonpathogenic E. coli and non-E.coli enteric and environmental bacterial strains. The results showed a high degree of specificity of the primers and probes for strains from corresponding virotypes and no reaction with the nontarget bacterial strains. The proposed multiplex PCR-DNA probing assay provides rapid and specific detection of four major virotypes of E. coli.
Leigh Watterworth; Edward Topp; Heidi Schraft; Kam Tin Leung
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of microbiological methods     Volume:  60     ISSN:  0167-7012     ISO Abbreviation:  J. Microbiol. Methods     Publication Date:  2005 Jan 
Date Detail:
Created Date:  2004-11-29     Completed Date:  2005-03-10     Revised Date:  2008-08-20    
Medline Journal Info:
Nlm Unique ID:  8306883     Medline TA:  J Microbiol Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  93-105     Citation Subset:  IM    
Department of Biology, Lakehead University, 955 Oliver Road, Thunder Bay, Ontario, Canada P7B 5E1.
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MeSH Terms
Adhesins, Bacterial / chemistry,  genetics
Bacterial Toxins / chemistry,  genetics
Chemiluminescent Measurements
DNA Probes / chemistry,  genetics
DNA, Bacterial / chemistry,  genetics
Enterotoxins / chemistry,  genetics
Escherichia coli / genetics*,  isolation & purification*,  pathogenicity
Escherichia coli Proteins / chemistry,  genetics
Plasmids / chemistry,  genetics
Polymerase Chain Reaction / methods*
Shiga Toxin 1 / chemistry,  genetics
Shiga Toxin 2 / chemistry,  genetics
Reg. No./Substance:
0/Adhesins, Bacterial; 0/Bacterial Toxins; 0/DNA Probes; 0/DNA, Bacterial; 0/Enterotoxins; 0/Escherichia coli Proteins; 0/Shiga Toxin 1; 0/Shiga Toxin 2; 0/heat stable toxin (E coli); 0/heat-labile enterotoxin, E coli

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