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A comparison of different DNA extraction methods and molecular techniques for the detection and identification of foodborne pathogens
Foodborne infections continue to plague Europe. Food safety monitoring is in crisis as the existing techniques for detecting pathogens do not keep up with the global rising of food production and consumption. Thus, the development of innovative techniques for detecting and identifying pathogenic bac...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
AIMS Press
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8500797/ https://www.ncbi.nlm.nih.gov/pubmed/34708174 http://dx.doi.org/10.3934/microbiol.2021019 |
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author | Papatheodorou, Spyridon Andreas Halvatsiotis, Panagiotis Houhoula, Dimitra |
author_facet | Papatheodorou, Spyridon Andreas Halvatsiotis, Panagiotis Houhoula, Dimitra |
author_sort | Papatheodorou, Spyridon Andreas |
collection | PubMed |
description | Foodborne infections continue to plague Europe. Food safety monitoring is in crisis as the existing techniques for detecting pathogens do not keep up with the global rising of food production and consumption. Thus, the development of innovative techniques for detecting and identifying pathogenic bacteria has become critical. The aim of the present study was firstly to develop an innovative simple and low cost method of extracting bacterial DNA from contaminated food and water samples with Salmonella enteric(a) subsp. enteric(a) serovar Typhimurium and Listeria monocytogenes and its comparison with two commercial DNA extraction kits (Qiagen, Macherey-Nagel). Finally, pathogens' detection using two molecular techniques (PCR-electrophoresis, LAMP), in order to evaluate the best combination of DNA extraction and identification based on their sensitivity, cost, rapidity and simplicity. Considering the above criteria, among them, best was proved an in-house bacterial DNA extraction method, based on the chloroform-isoamyl alcohol protocol, with certain modifications. This technique showed statistically similar results in terms of sensitivity, compared to the commercial kits, while at the same time maintained high rapidity and much lower cost. Lastly, between the molecular techniques, LAMP was found more promising considering its simplicity, high rapidity and sensitivity. Conclusively, the in-house DNA extraction method along with the LAMP technique, was proven to be the best among the presented combinations. |
format | Online Article Text |
id | pubmed-8500797 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | AIMS Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-85007972021-10-26 A comparison of different DNA extraction methods and molecular techniques for the detection and identification of foodborne pathogens Papatheodorou, Spyridon Andreas Halvatsiotis, Panagiotis Houhoula, Dimitra AIMS Microbiol Research Article Foodborne infections continue to plague Europe. Food safety monitoring is in crisis as the existing techniques for detecting pathogens do not keep up with the global rising of food production and consumption. Thus, the development of innovative techniques for detecting and identifying pathogenic bacteria has become critical. The aim of the present study was firstly to develop an innovative simple and low cost method of extracting bacterial DNA from contaminated food and water samples with Salmonella enteric(a) subsp. enteric(a) serovar Typhimurium and Listeria monocytogenes and its comparison with two commercial DNA extraction kits (Qiagen, Macherey-Nagel). Finally, pathogens' detection using two molecular techniques (PCR-electrophoresis, LAMP), in order to evaluate the best combination of DNA extraction and identification based on their sensitivity, cost, rapidity and simplicity. Considering the above criteria, among them, best was proved an in-house bacterial DNA extraction method, based on the chloroform-isoamyl alcohol protocol, with certain modifications. This technique showed statistically similar results in terms of sensitivity, compared to the commercial kits, while at the same time maintained high rapidity and much lower cost. Lastly, between the molecular techniques, LAMP was found more promising considering its simplicity, high rapidity and sensitivity. Conclusively, the in-house DNA extraction method along with the LAMP technique, was proven to be the best among the presented combinations. AIMS Press 2021-09-09 /pmc/articles/PMC8500797/ /pubmed/34708174 http://dx.doi.org/10.3934/microbiol.2021019 Text en © 2021 the Author(s), licensee AIMS Press https://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 (https://creativecommons.org/licenses/by/4.0/) ) |
spellingShingle | Research Article Papatheodorou, Spyridon Andreas Halvatsiotis, Panagiotis Houhoula, Dimitra A comparison of different DNA extraction methods and molecular techniques for the detection and identification of foodborne pathogens |
title | A comparison of different DNA extraction methods and molecular techniques for the detection and identification of foodborne pathogens |
title_full | A comparison of different DNA extraction methods and molecular techniques for the detection and identification of foodborne pathogens |
title_fullStr | A comparison of different DNA extraction methods and molecular techniques for the detection and identification of foodborne pathogens |
title_full_unstemmed | A comparison of different DNA extraction methods and molecular techniques for the detection and identification of foodborne pathogens |
title_short | A comparison of different DNA extraction methods and molecular techniques for the detection and identification of foodborne pathogens |
title_sort | comparison of different dna extraction methods and molecular techniques for the detection and identification of foodborne pathogens |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8500797/ https://www.ncbi.nlm.nih.gov/pubmed/34708174 http://dx.doi.org/10.3934/microbiol.2021019 |
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