Document Detail


Multiplex PCR-DNA probe assay for the detection of pathogenic Escherichia coli.
MedLine Citation:
PMID:  15567229     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Leigh Watterworth; Edward Topp; Heidi Schraft; Kam Tin Leung
Related Documents :
23587949 - Key role for efflux in the preservative susceptibility and adaptive resistance of burkh...
20134249 - Structure and diversity of arsenic resistant bacteria in an old tin mine area of thailand.
16232659 - Design of pcr primers and gene probes for the general detection of bacterial population...
18272709 - New immuno-pcr assay for detection of low concentrations of shiga toxin 2 and its varia...
11497219 - Epidemiological survey of orientia tsutsugamushi distribution in field rodents in saita...
9399509 - Identification of pathogenic leptospira genospecies by continuous monitoring of fluorog...
2440949 - C3b generation is affected by the structure of the o-antigen polysaccharide in lipopoly...
17395329 - Detection of campylobacter jejuni and campylobacter coli in chicken meat samples by rea...
22212129 - A pestivirus dna vaccine based on a non-antibiotic resistance escherichia coli essentia...
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    
Affiliation:
Department of Biology, Lakehead University, 955 Oliver Road, Thunder Bay, Ontario, Canada P7B 5E1.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
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
Chemical
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

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Capillary electrophoresis single-strand conformation polymorphism analysis for monitoring soil bacte...
Next Document:  McRAPD as a new approach to rapid and accurate identification of pathogenic yeasts.