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Improved sample treatment protocol for accurate detection of live Salmonella spp. in food samples by viability PCR

Culture-based detection is still considered as the standard way for detection of Salmonella in foods, although molecular methods, such as viability PCR (vPCR), have been introduced to overcome some disadvantages of traditional culture methods. Despite the success of the vPCR methodology, the problem...

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Autores principales: Dinh Thanh, Mai, Agustí, Gemma, Mader, Anneluise, Appel, Bernd, Codony, Francesc
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5726647/
https://www.ncbi.nlm.nih.gov/pubmed/29232387
http://dx.doi.org/10.1371/journal.pone.0189302
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author Dinh Thanh, Mai
Agustí, Gemma
Mader, Anneluise
Appel, Bernd
Codony, Francesc
author_facet Dinh Thanh, Mai
Agustí, Gemma
Mader, Anneluise
Appel, Bernd
Codony, Francesc
author_sort Dinh Thanh, Mai
collection PubMed
description Culture-based detection is still considered as the standard way for detection of Salmonella in foods, although molecular methods, such as viability PCR (vPCR), have been introduced to overcome some disadvantages of traditional culture methods. Despite the success of the vPCR methodology, the problem of false-positive results is a major drawback, especially when applied to environmental samples, hindering the interpretation of the results. To improve the efficiency of vPCR, many approaches have been introduced by several authors during the last years. In the present work, the combination of PEMAX dye, double tube change, and double photo-activation step was established as a strategy to improve vPCR protocol. By combining these approaches, we developed an improved sample treatment protocol able to neutralize DNA signals of up to 5.0×10(7) dead cells/sample from both pure culture and artificially contaminated food samples. Our results indicate that vPCR can work reliable and has a potential for high throughput detection of live Salmonella cells in food samples, minimizing false-positive signals.
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spelling pubmed-57266472017-12-22 Improved sample treatment protocol for accurate detection of live Salmonella spp. in food samples by viability PCR Dinh Thanh, Mai Agustí, Gemma Mader, Anneluise Appel, Bernd Codony, Francesc PLoS One Research Article Culture-based detection is still considered as the standard way for detection of Salmonella in foods, although molecular methods, such as viability PCR (vPCR), have been introduced to overcome some disadvantages of traditional culture methods. Despite the success of the vPCR methodology, the problem of false-positive results is a major drawback, especially when applied to environmental samples, hindering the interpretation of the results. To improve the efficiency of vPCR, many approaches have been introduced by several authors during the last years. In the present work, the combination of PEMAX dye, double tube change, and double photo-activation step was established as a strategy to improve vPCR protocol. By combining these approaches, we developed an improved sample treatment protocol able to neutralize DNA signals of up to 5.0×10(7) dead cells/sample from both pure culture and artificially contaminated food samples. Our results indicate that vPCR can work reliable and has a potential for high throughput detection of live Salmonella cells in food samples, minimizing false-positive signals. Public Library of Science 2017-12-12 /pmc/articles/PMC5726647/ /pubmed/29232387 http://dx.doi.org/10.1371/journal.pone.0189302 Text en © 2017 Dinh Thanh et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Dinh Thanh, Mai
Agustí, Gemma
Mader, Anneluise
Appel, Bernd
Codony, Francesc
Improved sample treatment protocol for accurate detection of live Salmonella spp. in food samples by viability PCR
title Improved sample treatment protocol for accurate detection of live Salmonella spp. in food samples by viability PCR
title_full Improved sample treatment protocol for accurate detection of live Salmonella spp. in food samples by viability PCR
title_fullStr Improved sample treatment protocol for accurate detection of live Salmonella spp. in food samples by viability PCR
title_full_unstemmed Improved sample treatment protocol for accurate detection of live Salmonella spp. in food samples by viability PCR
title_short Improved sample treatment protocol for accurate detection of live Salmonella spp. in food samples by viability PCR
title_sort improved sample treatment protocol for accurate detection of live salmonella spp. in food samples by viability pcr
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5726647/
https://www.ncbi.nlm.nih.gov/pubmed/29232387
http://dx.doi.org/10.1371/journal.pone.0189302
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