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Improvement and validation of RAPD in combination with PFGE analysis of Salmonella enterica ssp enterica serovar Senftenberg strains isolated from feed mills

Löfström, Charlotta LU ; Eriksson, John LU ; Aspan, A ; Haggblom, P ; Gunnarsson, A ; Borch, E and Rådström, Peter LU (2006) In Veterinary Microbiology 114(3-4). p.345-351
Abstract
In 1995 and 1996 a Swedish feed mill had problems due to a persistent contamination of Salmonella enterica spp. enterica serovar Senftenberg that was difficult to eliminate. Forty-eight strains isolated from the feed mill, together with unrelated strains included to evaluate the discriminatory power and reproducibility, were analysed by pulsed-field gel electrophoresis (PFGE). The source of contamination in the feed mill was identified and preventative measures were taken, that led to a resolution of the problem. A previously developed randomly amplified polymorphic DNA (RAPD) protocol was used, to evaluate a rapid and low-cost alternative to PFGE typing. The use of the alternative thermostable DNA polymerase Tth was shown to increase the... (More)
In 1995 and 1996 a Swedish feed mill had problems due to a persistent contamination of Salmonella enterica spp. enterica serovar Senftenberg that was difficult to eliminate. Forty-eight strains isolated from the feed mill, together with unrelated strains included to evaluate the discriminatory power and reproducibility, were analysed by pulsed-field gel electrophoresis (PFGE). The source of contamination in the feed mill was identified and preventative measures were taken, that led to a resolution of the problem. A previously developed randomly amplified polymorphic DNA (RAPD) protocol was used, to evaluate a rapid and low-cost alternative to PFGE typing. The use of the alternative thermostable DNA polymerase Tth was shown to increase the reproducibility of the RAPD analysis. The reproducibility, in terms of Pearson's and Dice's similarity coefficients for duplicate runs, increased from 72.0 +/- 16.9% and 72.3 +/- 12.9% for Taq to 91.6 +/- 7.5% and 90.9 +/- 5.3% for the fingerprints obtained for the RAPD method employing Tth DNA polymerase. Simpson's index of diversity was calculated and found to be 0.580 for RAPD and 0.896 for PFGE. All of the seven RAPD, types could be subdivided into one or more PFGE types, whereas none of the 22 PFGE types was divided into more than one RAPD type. RAPD provides a simple, rapid and powerful screening method that can be used to initially select isolates for further analysis by PFGE (Less)
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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Salmonella, amplified polymorphic DNA, randomly, pulsed-field gel electrophoresis, animal feed, genotyping
in
Veterinary Microbiology
volume
114
issue
3-4
pages
345 - 351
publisher
Elsevier
external identifiers
  • pmid:16427217
  • wos:000237667400020
  • scopus:33646150925
  • pmid:16427217
ISSN
0378-1135
DOI
10.1016/j.vetmic.2005.12.015
language
English
LU publication?
yes
id
883ddb30-1920-4948-84de-c6a416fec5cc (old id 408782)
date added to LUP
2016-04-01 16:08:58
date last changed
2021-09-22 02:13:15
@article{883ddb30-1920-4948-84de-c6a416fec5cc,
  abstract     = {In 1995 and 1996 a Swedish feed mill had problems due to a persistent contamination of Salmonella enterica spp. enterica serovar Senftenberg that was difficult to eliminate. Forty-eight strains isolated from the feed mill, together with unrelated strains included to evaluate the discriminatory power and reproducibility, were analysed by pulsed-field gel electrophoresis (PFGE). The source of contamination in the feed mill was identified and preventative measures were taken, that led to a resolution of the problem. A previously developed randomly amplified polymorphic DNA (RAPD) protocol was used, to evaluate a rapid and low-cost alternative to PFGE typing. The use of the alternative thermostable DNA polymerase Tth was shown to increase the reproducibility of the RAPD analysis. The reproducibility, in terms of Pearson's and Dice's similarity coefficients for duplicate runs, increased from 72.0 +/- 16.9% and 72.3 +/- 12.9% for Taq to 91.6 +/- 7.5% and 90.9 +/- 5.3% for the fingerprints obtained for the RAPD method employing Tth DNA polymerase. Simpson's index of diversity was calculated and found to be 0.580 for RAPD and 0.896 for PFGE. All of the seven RAPD, types could be subdivided into one or more PFGE types, whereas none of the 22 PFGE types was divided into more than one RAPD type. RAPD provides a simple, rapid and powerful screening method that can be used to initially select isolates for further analysis by PFGE},
  author       = {Löfström, Charlotta and Eriksson, John and Aspan, A and Haggblom, P and Gunnarsson, A and Borch, E and Rådström, Peter},
  issn         = {0378-1135},
  language     = {eng},
  number       = {3-4},
  pages        = {345--351},
  publisher    = {Elsevier},
  series       = {Veterinary Microbiology},
  title        = {Improvement and validation of RAPD in combination with PFGE analysis of Salmonella enterica ssp enterica serovar Senftenberg strains isolated from feed mills},
  url          = {http://dx.doi.org/10.1016/j.vetmic.2005.12.015},
  doi          = {10.1016/j.vetmic.2005.12.015},
  volume       = {114},
  year         = {2006},
}