Detection of Food Spoilage and Pathogenic Bacteria Based on Ligation Detection Reaction Coupled to Flow-Through Hybridization on Membranes
Joint Authors
Barros-Velázquez, Jorge
Cremonesi, P.
Calo-Mata, Pilar
Castiglioni, B.
Fernández-No, Inmaculada
Böhme, Karola
Villa, Tomás G.
Severgnini, M.
Source
Issue
Vol. 2014, Issue 2014 (31 Dec. 2014), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2014-04-10
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Abstract EN
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.
American Psychological Association (APA)
Böhme, Karola& Cremonesi, P.& Severgnini, M.& Villa, Tomás G.& Fernández-No, Inmaculada& Barros-Velázquez, Jorge…[et al.]. 2014. Detection of Food Spoilage and Pathogenic Bacteria Based on Ligation Detection Reaction Coupled to Flow-Through Hybridization on Membranes. BioMed Research International،Vol. 2014, no. 2014, pp.1-11.
https://search.emarefa.net/detail/BIM-450262
Modern Language Association (MLA)
Böhme, Karola…[et al.]. Detection of Food Spoilage and Pathogenic Bacteria Based on Ligation Detection Reaction Coupled to Flow-Through Hybridization on Membranes. BioMed Research International No. 2014 (2014), pp.1-11.
https://search.emarefa.net/detail/BIM-450262
American Medical Association (AMA)
Böhme, Karola& Cremonesi, P.& Severgnini, M.& Villa, Tomás G.& Fernández-No, Inmaculada& Barros-Velázquez, Jorge…[et al.]. Detection of Food Spoilage and Pathogenic Bacteria Based on Ligation Detection Reaction Coupled to Flow-Through Hybridization on Membranes. BioMed Research International. 2014. Vol. 2014, no. 2014, pp.1-11.
https://search.emarefa.net/detail/BIM-450262
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-450262