Document Detail


Development of a PCR microplate-capture hybridization method for simple, fast and sensitive detection of Salmonella serovars in food.
MedLine Citation:
PMID:  9727199     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The authors have developed an easy and rapid detection and identification system for Salmonella spp. in food. The gene inv A was selected as the target sequence. Oligonucleotides derived from conserved regions of this gene were able to exclusively prime the amplification of a 389 bp fragment when Salmonella spp. DNA was used as the template. An internal Salmonella spp. specific DNA probe was used for confirmation of the amplified polymerase chain reaction(PCR)product, by Southern blot or microplate-capture hybridization assay. In this fashion the sensitivity of the method was increased 100-fold (4.5 fg total DNA). To validate the method, a total of 75 food samples were tested. The PCR-microplate capture hybridization assay is easy to perform and much faster than traditional detection methods for Salmonella spp. in food. Hybridization in microtitre plates is more readily observed than in Southern blot and is more sensitive than conventional agarose gel electrophoresis.
Authors:
M Manzano; L Cocolin; G Astori; C Pipan; G A Botta; C Cantoni; G Comi
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Publication Detail:
Type:  Comparative Study; Journal Article    
Journal Detail:
Title:  Molecular and cellular probes     Volume:  12     ISSN:  0890-8508     ISO Abbreviation:  Mol. Cell. Probes     Publication Date:  1998 Aug 
Date Detail:
Created Date:  1998-11-24     Completed Date:  1998-11-24     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8709751     Medline TA:  Mol Cell Probes     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  227-34     Citation Subset:  IM    
Affiliation:
Dipartimento di Scienze degli Alimenti, Facoltà di Agraria, Università di Udine, Italy.
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MeSH Terms
Descriptor/Qualifier:
Biotin
Blotting, Southern / methods
DNA, Complementary / chemical synthesis
Food Microbiology*
Nucleic Acid Hybridization / methods
Polymerase Chain Reaction / instrumentation,  methods*
Reproducibility of Results
Salmonella / genetics*,  isolation & purification*
Sensitivity and Specificity
Streptavidin
Chemical
Reg. No./Substance:
0/DNA, Complementary; 58-85-5/Biotin; 9013-20-1/Streptavidin

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


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