Document Detail


Magnetic bead hybridization to detect enterotoxigenic Escherichia coli strains associated with cattle in environmental water sources.
MedLine Citation:
PMID:  14569293     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A magnetic capture hybridization - polymerase chain reaction (MCH-PCR) method was used to increase the detection sensitivity of the enterotoxin gene LTIIa, used as a biomarker for waste in environmental samples. The samples were collected from cow lagoons of different farms and from environmental waters. Total DNA was extracted from colonies grown on mTEC medium or directly from environmental samples. The cow-specific Escherichia coli LTIIa gene was used as a DNA marker. A LTIIa-specific oligonucleotide probe was designed to capture the LTIIa marker during the MCH, followed by PCR. Varying levels of humic acid were added to the DNA extracts to evaluate the sensitivity and effectiveness of MCH-PCR. The minimal detection limit of MCH-PCR for the LTIIa gene was 2.5 ag/muL DNA. In the presence of humic acid, MCH-PCR was able to increase the detection sensitivity 10 000-fold over that of conventional PCR. The MCH-PCR could also detect one cell with the LTIIa DNA marker in a 1-L seeded environmental water sample. Results in this study indicate that MCH-PCR is more sensitive than nested PCR in testing environmental samples.
Authors:
Yu-Li Tsai; Joanne Y Le; Betty H Olson
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Canadian journal of microbiology     Volume:  49     ISSN:  0008-4166     ISO Abbreviation:  Can. J. Microbiol.     Publication Date:  2003 Jun 
Date Detail:
Created Date:  2003-10-21     Completed Date:  2003-12-05     Revised Date:  2008-08-20    
Medline Journal Info:
Nlm Unique ID:  0372707     Medline TA:  Can J Microbiol     Country:  Canada    
Other Details:
Languages:  eng     Pagination:  391-8     Citation Subset:  IM    
Affiliation:
Department of Environmental Health Science and Policy, University of California, Irvine 92697, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Bacterial Toxins / genetics*
Cattle / microbiology*
Enterotoxins / genetics*
Escherichia coli / isolation & purification*
Escherichia coli Proteins*
Feces / microbiology*
Genes, Bacterial
Humic Substances
Magnetics
Microspheres
Nucleic Acid Hybridization / methods*
Polymerase Chain Reaction / methods*
Sensitivity and Specificity
Water Microbiology*
Water Pollution / analysis
Chemical
Reg. No./Substance:
0/Bacterial Toxins; 0/Enterotoxins; 0/Escherichia coli Proteins; 0/Humic Substances; 0/heat-labile enterotoxin, E coli

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


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