Electrochemical genosensing of Salmonella, Listeria and Escherichia coli on silica magnetic particles
Autor: | María Isabel Pividori, Pilar Cortés, Delfina Brandão, Salvador Alegret, Susana Liébana, Susana Campoy |
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Rok vydání: | 2016 |
Předmět: |
Salmonella
Listeria 02 engineering and technology medicine.disease_cause Polymerase Chain Reaction 01 natural sciences Biochemistry Analytical Chemistry Magnetics chemistry.chemical_compound Guanidinium thiocyanate Listeria monocytogenes Escherichia coli medicine Environmental Chemistry Digoxigenin Spectroscopy Chromatography biology 010401 analytical chemistry Silicon Dioxide 021001 nanoscience & nanotechnology biology.organism_classification Molecular biology 0104 chemical sciences chemistry Salmonella enterica Microscopy Electron Scanning Nucleic acid 0210 nano-technology |
Zdroj: | Analytica Chimica Acta. 904:1-9 |
ISSN: | 0003-2670 |
Popis: | A magneto-genosensing approach for the detection of the three most common pathogenic bacteria in food safety, such as Salmonella, Listeria and Escherichia coli is presented. The methodology is based on the detection of the tagged amplified DNA obtained by single-tagging PCR with a set of specific primers for each pathogen, followed by electrochemical magneto-genosensing on silica magnetic particles. A set of primers were selected for the amplification of the invA (278 bp), prfA (217 bp) and eaeA (151 bp) being one of the primers for each set tagged with fluorescein, biotin and digoxigenin coding for Salmonella enterica, Listeria monocytogenes and E. coli, respectively. The single-tagged amplicons were then immobilized on silica MPs based on the nucleic acid-binding properties of silica particles in the presence of the chaotropic agent as guanidinium thiocyanate. The assessment of the silica MPs as a platform for electrochemical magneto-genosensing is described, including the main parameters to selectively attach longer dsDNA fragments instead of shorter ssDNA primers based on their negative charge density of the sugar-phosphate backbone. This approach resulted to be a promising detection tool with sensing features of rapidity and sensitivity very suitable to be implemented on DNA biosensors and microfluidic platforms. |
Databáze: | OpenAIRE |
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