Document Detail

Detection of low numbers of Salmonella in environmental water, sewage and food samples by a nested polymerase chain reaction assay.
MedLine Citation:
PMID:  10540245     Owner:  NLM     Status:  MEDLINE    
A polymerase chain reaction (PCR) assay with two nested pairs of primers selected from conserved sequences within a 2.3 kb randomly cloned DNA fragment from the Salmonella typhimurium chromosome was developed. The nested PCR assay correctly identified 128 of a total of 129 Salmonella strains belonging to subspecies I, II, IIIb and IV. One strain of Salm. arizona (ssp. IIIa) tested negative. No PCR products were obtained from any of the 31 non-Salmonella strains examined. The sensitivity of the assay was 2 cfu, as determined by analysis of proteinase K-treated boiled lysates of Salm. typhimurium. The performance of the assay was evaluated for environmental water, sewage and food samples spiked with Salm. typhimurium. Water and sewage samples were filtered and filters were enriched overnight in a non-selective medium. Prior to PCR, the broth cultures were subjected to a rapid and simple preparation procedure consisting of centrifugation, proteinase K treatment and boiling. This assay enabled detection of 10 cfu 100 ml(-1) water with background levels of up to 8700 heterotrophic organisms ml(-1) and 10000 cfu of coliform organisms 100 ml(-1) water. Spiked food samples were analysed with and without overnight enrichment in a non-selective medium using the same assay as above. Nested PCR performed on enriched broths enabled detection of <10 cfu g(-1) food. Variable results were obtained for food samples examined without prior enrichment and most results were negative. This rapid and simple assay provides a sensitive and specific means of screening drinking water or environmental water samples, as well as food samples, for the presence of Salmonella spp.
A S Waage; T Vardund; V Lund; G Kapperud
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of applied microbiology     Volume:  87     ISSN:  1364-5072     ISO Abbreviation:  J. Appl. Microbiol.     Publication Date:  1999 Sep 
Date Detail:
Created Date:  1999-11-22     Completed Date:  1999-11-22     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9706280     Medline TA:  J Appl Microbiol     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  418-28     Citation Subset:  IM    
Department of Bacteriology, National Institute of Public Health, Norwegian College of Veterinary Medicine, Oslo, Norway.
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MeSH Terms
DNA Primers
Food Microbiology*
Polymerase Chain Reaction / methods*
Salmonella / genetics,  isolation & purification*
Sensitivity and Specificity
Sewage / microbiology*
Water Microbiology*
Reg. No./Substance:
0/DNA Primers; 0/Sewage

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