RT Journal Article
T1 Detection of food spoilage and pathogenic bacteria based on ligation detection reaction coupled to flow-through hybridization on membranes
A1 Böhme, Karola
A1 Cremonesi, P.
A1 Severgnini, M.
A1 González Villa, Tomás
A1 Fernández No, Inmaculada Concepción
A1 Barros Velázquez, Jorge
A1 Castiglioni, B.
A1 Calo Mata, María Pilar
K1 Food spoilage
K1 Membranes K
AB 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
PB Hindawi
SN 2314-6133
YR 2014
FD 2014
LK http://hdl.handle.net/10347/21591
UL http://hdl.handle.net/10347/21591
LA eng
NO 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
NO This work was funded by the Project 10PXIB261045PR from Xunta de Galicia and by the Projects AGL2010-19646 from the Spanish Ministry of Science and Technology. The predoctoralresearch stay of K. Böhme in Italywas funded by a FEMS Research Fellowship
DS Minerva
RD 3 may 2026