title: Food sensing: selection and characterization of DNA aptamers to Alicyclobacillus spores for trapping and detection from orange juice.
abstract: The quality of the beverage industry's products has to be constantly monitored to fulfill consumers' high expectations. The thermo-acidophilic Gram-positive Alicyclobacillus spp. are not pathogenic, but their heat-resistant endospores can survive juice-processing conditions and have become a major economic concern for the fruit juice industry. Current detection methods rely on cultivation, isolation, and organism identification, which can take up to a week, resulting in economic loss. This work presents the selection and identification of DNA aptamers targeting Alicyclobacillus spores by spore-SELEX (systematic evolution of ligands by exponential enrichment) in orange-juice-simulating buffer. The selection process was verified by various techniques, including flow cytometric binding assays, radioactive binding assays, and agarose gel electrophoresis. The subsequent aptamer characterization included the determination of dissociations constants and selectivity by different techniques, such as surface plasmon resonance spectroscopy and fluorescence microscopy. In summary, 10 different aptamers with an affinity to Alicyclobacillus spp. have been developed, analyzed, and characterized in terms of affinity and specificity.
authors: Hamburg School of Food Science, Institute of Food Chemistry, University of Hamburg , Grindelallee 117, 20146 Hamburg, Germany.
authors: Tim
authors: T
authors: Hünniger
authors: Christin
authors: C
authors: Fischer
authors: Hauke
authors: H
authors: Wessels
authors: Antonia
authors: A
authors: Hoffmann
authors: Angelika
authors: A
authors: Paschke-Kratzin
authors: Ilka
authors: I
authors: Haase
authors: Markus
authors: M
authors: Fischer
mesh_terms: D056450:Alicyclobacillus / Q000235:genetics / Q000302:isolation & purification*; D052157:Aptamers, Nucleotide / Q000235:genetics; D001628:Beverages / Q000032:analysis / Q000382:microbiology*; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D032084:Citrus sinensis / Q000382:microbiology*; D005506:Food Contamination / Q000032:analysis; D052156:SELEX Aptamer Technique / Q000295:instrumentation / Q000379:methods*; D013171:Spores, Bacterial / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D052157:Aptamers, Nucleotide
keywords: Alicyclobacillus; FACS; SELEX; SPR; aptamers; enrichment; juice; selection; spores