Detection of food spoilage and pathogenic bacteria based on ligation detection reaction coupled to flow-through hybridization on membranes
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Título: | Detection of food spoilage and pathogenic bacteria based on ligation detection reaction coupled to flow-through hybridization on membranes |
Autor/a: | Böhme, Karola Cremonesi, P. Severgnini, M. González Villa, Tomás Fernández No, Inmaculada Concepción Barros Velázquez, Jorge Castiglioni, B. Calo Mata, María del Pilar |
Centro/Departamento: | Universidade de Santiago de Compostela. Departamento de Química Analítica, Nutrición e Bromatoloxía |
Palabras chave: | Food spoilage | Membranes K | |
Data: | 2014 |
Editor: | Hindawi |
Cita bibliográfica: | Böhme, K, Cremonesi, P., Severgnini, M., Villa,Tomas G., Fernández No, I.C.,Barros Velázquez, J., Castiglioni, B., Calo Mata,P. (2013). Detection of food spoilage and pathogenic bacteria based on ligation detection reaction coupled to flow-through hybridization on membranes. " Biomed Research International", vol. 2014 |
Resumo: | Traditional culturing methods are still commonly applied for bacterial identification in the food control sector, despite being time and labor intensive. Microarray technologies represent an interesting alternative. However, they require higher costs and technical expertise, making them still inappropriate for microbial routine analysis. The present study describes the development of an efficient method for bacterial identification based on flow-through reverse dot-blot (FT-RDB) hybridization on membranes, coupled to the high specific ligation detection reaction (LDR). First, the methodology was optimized by testing different types of ligase enzymes, labeling, and membranes. Furthermore, specific oligonucleotide probes were designed based on the 16S rRNA gene, using the bioinformatic tool Oligonucleotide Retrieving for Molecular Applications (ORMA). Four probes were selected and synthesized, being specific for Aeromonas spp., Pseudomonas spp., Shewanella spp., and Morganella morganii, respectively. For the validation of the probes, 16 reference strains from type culture collections were tested by LDR and FT-RDB hybridization using universal arrays spotted onto membranes. In conclusion, the described methodology could be applied for the rapid, accurate, and cost-effective identification of bacterial species, exhibiting special relevance in food safety and quality |
Versión do editor: | https://doi.org/10.1155/2014/156323 |
URI: | http://hdl.handle.net/10347/21591 |
DOI: | 10.1155/2014/156323 |
ISSN: | 2314-6133 |
E-ISSN: | 2314-6141 |
Dereitos: | Copyright © 2014 K. Böhme et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited |
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