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Clostridium
Published in Dongyou Liu, Handbook of Foodborne Diseases, 2018
Emilio Aranda, Mar Rodríguez, María G. Córdoba, María J. Benito, Juan J. Córdoba
ELISA and immuno-PCR and ELISA based on protein antibody microarray have been recently reported to detect BoNTs A, B, C, D, E, and F in complex clinical, food, and environmental samples at higher sensitivity than mouse bioassay.115,116 A number of rapid affinity immunochromatography column (AICC) assays are also described for the detection of BoNT serotypes A, B, E, and F in food matrices. The reported detection limit of 0.5 ng A BoNT is twofold more sensitive than that of earlier lateral flow methods. For serotypes B, E, and F, the minimum detection limit ranged from 5 to 50 ng. Although not as sensitive as ELISA or mouse bioassays, immunochromatographic methods generally are rapid, requiring only 15–30 minutes to complete, without enrichment steps, making them highly amenable to use in the field.
Clostridium
Published in Dongyou Liu, Laboratory Models for Foodborne Infections, 2017
Emilio Aranda, María G. Córdoba, María J. Benito, Juan José Córdoba
Being the most widely distributed pathogenic species within the Clostridium genus, C. botulinum, C. perfringens, and C. difficile are frequently involved in foodborne outbreaks. Accurate methods for isolating and detecting these pathogenic clostridia in raw material and processed foods are necessary to take corrective measures throughout food processing that reduce potential clostridial cases. For isolation, detection, and characterization of C. botulinum, C. perfringens, and C. difficile from food samples, traditional microbiological culture techniques and molecular methods (PCR and qPCR) are available. When these methods are used in foodborne investigation, confirmation of toxin production by rapid methods such as ELISA, immuno-PCR, or ELISA based on protein antibody microarray is important. While these methods reveal the presence of pathogenic Clostridium species and/or the corresponding toxins in food, no information is available about the effect of these microorganisms in live cells, which is required in most of the cases during clostridial foodborne investigation. Bioassays using laboratory animal or cells culture methods help determine the effects of neurotoxin or enterotoxins in live organisms. Based on the ability of a suspected food extracts to induce symptoms in laboratory animal models and/or superantigenic action in cell culture models, these methods offer a very valuable alternative for foodborne clostridia investigation. The mouse neutralization bioassay, using either monovalent, toxin-type-specific botulinum antitoxin or polyvalent antibotulinum antitoxin is the most valuable method for foodborne botulism investigation. Rabbit intestinal loops, rat, and mice laboratory models are available for C. perfringens foodborne investigation. For C. difficile investigation, hamster, mouse, and pigs may be utilized. In addition, cell lines provide an alternative to laboratory animal models for foodborne clostridial cases investigation. For example, rat spinal cord, chick embryo neuronal, and neuroblastoma cell lines are available for accurate investigation of foodborne botulism. For the study of pathogenic C. perfringens and C. difficile, human intestinal epithelial lines such as Caco-2 cells have been reported as sensitive method.
Emerging proteomic biomarkers of X-linked muscular dystrophy
Published in Expert Review of Molecular Diagnostics, 2019
Paul Dowling, Sandra Murphy, Margit Zweyer, Maren Raucamp, Dieter Swandulla, Kay Ohlendieck
The typical workflow of an antibody microarray involves the choice of an appropriate antibody array platform (solid or bead), blocking of non-specific reactive groups, selection of suitable samples containing soluble proteins of interest and incubation with the antibody array, where targeted proteins from the sample are captured by the antibodies. The resulting binding events are reported precisely by fluorescent labelling of the sample or by the addition of a secondary detection reagent. Most of the antibody arrays can achieve pg/ml detection for specific antigens, similar to single ELISA detection methodology [90]. However, to overcome potential cross-reactivity and target protein abundance issues and concerns at detecting low/high abundant target proteins on the same array, more specialised arrays can be developed to mitigate for these trepidations. Current off-the-shelf antibody array panels include those specifically developed to detect and quantitate various cytokines, chemokines, adipokines, growth factors, proteases, soluble receptors and adhesion molecules. The custom option offers researchers the flexibility to design tailored panels to meet niche demands where novel biomarkers or marker molecules harbouring specific post-translation modifications need to be included as part of an antibody array panel.
Exploring blood-brain barrier hyperpermeability and potential biomarkers in traumatic brain injury
Published in Baylor University Medical Center Proceedings, 2020
Bobby Darnell Robinson, Binu Tharakan, Angela Lomas, Katie Wiggins-Dohlvik, Himakarnika Alluri, Chinchusha Anasooya Shaji, Daniel Jupiter, Claire Larson Isbell
Collected blood samples from animals were shipped to RayBiotech, Inc. (Norcross, GA) for antibody microarray analysis to screen for potential biomarkers. The following analytes were studied: ZO-1, junctional adhesion molecule-1 (JAM-1), tricellulin, occludin, vascular endothelial (VE)-cadherin, β-catenin, caspase-3, caspase-8, B-cell lymphoma 2 (Bcl-2)–associated X (BAX), Bcl-2–associated death promoter (BAD), cytochrome C, Bcl-2 homologue-3 interacting-domain death agonist (BID), Bcl-2-like protein 11 (BIM), soluble Fas protein (sFas), Matrix metalloproteinase-9 (MMP-9), interleukin (IL)−1β, and IL-17.