BIOMAT Database Logo BIOMAT Database Logo

Rapid and specific detection of Salmonella infections using chemically modified nucleic acid probes. 2019

Isabel Machado, and Victoria Garrido, and Luiza I Hernandez, and Juliana Botero, and Nora Bastida, and Beatriz San-Roman, and María-Jesús Grilló, and Frank J Hernandez
SOMAprobes, Science and Technology Park of Gipuzkoa, Donostia-San Sebastian, 20009, Spain.

Salmonella is a leading source of bacterial foodborne illness in humans, causing gastroenteritis outbreaks with bacteraemia occurrences that can lead to clinical complications and death. Eggs, poultry and pig products are considered as the main carriers of the pathogenic Salmonella for humans. To prevent this relevant zoonosis, key changes in food safety regulations were undertaken to improve controls in the food production chain. Despite these measures, large outbreaks of salmonellosis were reported worldwide in the last decade. Thus, new strategies for Salmonella detection are a priority for both, food safety and public health authorities. Such detection systems should provide significant reduction in diagnostic time (hours) compared to the currently available methods (days). Herein, we report on the discovery and characterization of nucleic acid probes for the sensitive and specific detection of live Salmonella within less than 8 h of incubation. We are the first to postulate the nuclease activity derived from Salmonella as biomarker of infection and its utility to develop innovative detection strategies. Our results have shown the screening and identification of two oligonucleotide sequences (substrates) as the most promising probes for detecting Salmonella - Sal-3 and Sal-5. The detection limits for both probes were determined with the reference Salmonella Typhimurium (STM 1) and Salmonella Enteritidis (SE 1) cultures. Sal-3 has reported LOD values around 105 CFU mL-1 for STM 1 and 104 CFU mL-1 for SE 1, while Sal-5 proves to be a slightly better probe, with LODs of 104 CFU mL-1 for STM 1 and 104 CFU mL-1 for SE 1. Both selected probes have shown the capability to recognize 49 out of 51 different Salmonella serotypes tested in vitro and the most frequent serotypes in porcine mesenteric lymph nodes as a standard sample used in fattening-pig salmonellosis baseline studies. Notably, our results showed 100% correlation between nuclease detection and the PCR-InvA or ISO-6579 standard method, underlining the great potential of this innovative nucleic acids technology to be implemented as a rapid method for food safety testing.

UI MeSH Term Description Entries
D008958 Models, Molecular Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures. Molecular Models,Model, Molecular,Molecular Model
D009690 Nucleic Acid Conformation The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape. DNA Conformation,RNA Conformation,Conformation, DNA,Conformation, Nucleic Acid,Conformation, RNA,Conformations, DNA,Conformations, Nucleic Acid,Conformations, RNA,DNA Conformations,Nucleic Acid Conformations,RNA Conformations
D011487 Protein Conformation The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain). Conformation, Protein,Conformations, Protein,Protein Conformations
D003851 Deoxyribonucleases Enzymes which catalyze the hydrolases of ester bonds within DNA. EC 3.1.-. DNAase,DNase,Deoxyribonuclease,Desoxyribonuclease,Desoxyribonucleases,Nucleases, DNA,Acid DNase,Alkaline DNase,DNA Nucleases,DNase, Acid,DNase, Alkaline
D005516 Food Microbiology The presence of bacteria, viruses, and fungi in food and food products. This term is not restricted to pathogenic organisms: the presence of various non-pathogenic bacteria and fungi in cheeses and wines, for example, is included in this concept. Microbiology, Food
D012475 Salmonella A genus of gram-negative, facultatively anaerobic, rod-shaped bacteria that utilizes citrate as a sole carbon source. It is pathogenic for humans, causing enteric fevers, gastroenteritis, and bacteremia. Food poisoning is the most common clinical manifestation. Organisms within this genus are separated on the basis of antigenic characteristics, sugar fermentation patterns, and bacteriophage susceptibility.
D013997 Time Factors Elements of limited time intervals, contributing to particular results or situations. Time Series,Factor, Time,Time Factor
D015345 Oligonucleotide Probes Synthetic or natural oligonucleotides used in hybridization studies in order to identify and study specific nucleic acid fragments, e.g., DNA segments near or within a specific gene locus or gene. The probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the probe include the radioisotope labels 32P and 125I and the chemical label biotin. Oligodeoxyribonucleotide Probes,Oligonucleotide Probe,Oligoribonucleotide Probes,Probe, Oligonucleotide,Probes, Oligodeoxyribonucleotide,Probes, Oligonucleotide,Probes, Oligoribonucleotide
D057230 Limit of Detection Concentration or quantity that is derived from the smallest measure that can be detected with reasonable certainty for a given analytical procedure. Limits of Detection,Detection Limit,Detection Limits
D059022 Food Safety Activities involved in ensuring the safety of FOOD including avoidance of bacterial and other contamination. Safety, Food

Related Publications

Isabel Machado, and Victoria Garrido, and Luiza I Hernandez, and Juliana Botero, and Nora Bastida, and Beatriz San-Roman, and María-Jesús Grilló, and Frank J Hernandez
Specific nucleic acid detection using photophysical properties of quantum dot probes.
February 2010, Analytical chemistry,
Isabel Machado, and Victoria Garrido, and Luiza I Hernandez, and Juliana Botero, and Nora Bastida, and Beatriz San-Roman, and María-Jesús Grilló, and Frank J Hernandez
2-Acetylaminofluorene-modified probes for the indirect hybridocytochemical detection of specific nucleic acid sequences.
July 1984, Experimental cell research,
Isabel Machado, and Victoria Garrido, and Luiza I Hernandez, and Juliana Botero, and Nora Bastida, and Beatriz San-Roman, and María-Jesús Grilló, and Frank J Hernandez
Detection and quantification of genetically modified organisms using very short, locked nucleic acid TaqMan probes.
June 2008, Journal of agricultural and food chemistry,
Isabel Machado, and Victoria Garrido, and Luiza I Hernandez, and Juliana Botero, and Nora Bastida, and Beatriz San-Roman, and María-Jesús Grilló, and Frank J Hernandez
Artificial specific binders directly recovered from chemically modified nucleic acid libraries.
January 2012, Journal of nucleic acids,
Isabel Machado, and Victoria Garrido, and Luiza I Hernandez, and Juliana Botero, and Nora Bastida, and Beatriz San-Roman, and María-Jesús Grilló, and Frank J Hernandez
Chemically modified nucleic acids as immunodetectable probes in hybridization experiments.
June 1984, Proceedings of the National Academy of Sciences of the United States of America,
Isabel Machado, and Victoria Garrido, and Luiza I Hernandez, and Juliana Botero, and Nora Bastida, and Beatriz San-Roman, and María-Jesús Grilló, and Frank J Hernandez
Crosslinker-modified nucleic acid probes for improved target identification and biomarker detection.
April 2021, RSC chemical biology,
Isabel Machado, and Victoria Garrido, and Luiza I Hernandez, and Juliana Botero, and Nora Bastida, and Beatriz San-Roman, and María-Jesús Grilló, and Frank J Hernandez
Rapid detection, identification, and enumeration of Pseudomonas aeruginosa in bottled water using peptide nucleic acid probes.
November 2000, Journal of microbiological methods,
Isabel Machado, and Victoria Garrido, and Luiza I Hernandez, and Juliana Botero, and Nora Bastida, and Beatriz San-Roman, and María-Jesús Grilló, and Frank J Hernandez
Focused upon hybridization: rapid and high sensitivity detection of DNA using isotachophoresis and peptide nucleic acid probes.
September 2015, Analytical chemistry,
Isabel Machado, and Victoria Garrido, and Luiza I Hernandez, and Juliana Botero, and Nora Bastida, and Beatriz San-Roman, and María-Jesús Grilló, and Frank J Hernandez
MicroRNA detection by northern blotting using locked nucleic acid probes.
January 2008, Nature protocols,
Isabel Machado, and Victoria Garrido, and Luiza I Hernandez, and Juliana Botero, and Nora Bastida, and Beatriz San-Roman, and María-Jesús Grilló, and Frank J Hernandez
Fluorescence in situ Hybridization method using Peptide Nucleic Acid probes for rapid detection of Lactobacillus and Gardnerella spp.
April 2013, BMC microbiology,
Copied contents to your clipboard!