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Authors = Jeffery McGarvey

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9 pages, 2100 KiB  
Article
Rapid Microfluidic Assay for the Detection of Botulinum Neurotoxin in Animal Sera
by Lmar Babrak, Alice Lin, Larry H. Stanker, Jeffery McGarvey and Robert Hnasko
Toxins 2016, 8(1), 13; https://doi.org/10.3390/toxins8010013 - 4 Jan 2016
Cited by 22 | Viewed by 7036
Abstract
Potent Botulinum neurotoxins (BoNTs) represent a threat to public health and safety. Botulism is a disease caused by BoNT intoxication that results in muscle paralysis that can be fatal. Sensitive assays capable of detecting BoNTs from different substrates and settings are essential to [...] Read more.
Potent Botulinum neurotoxins (BoNTs) represent a threat to public health and safety. Botulism is a disease caused by BoNT intoxication that results in muscle paralysis that can be fatal. Sensitive assays capable of detecting BoNTs from different substrates and settings are essential to limit foodborne contamination and morbidity. In this report, we describe a rapid 96-well microfluidic double sandwich immunoassay for the sensitive detection of BoNT-A from animal sera. This BoNT microfluidic assay requires only 5 μL of serum, provides results in 75 min using a standard fluorescence microplate reader and generates minimal hazardous waste. The assay has a <30 pg·mL−1 limit of detection (LOD) of BoNT-A from spiked human serum. This sensitive microfluidic BoNT-A assay offers a fast and simplified workflow suitable for the detection of BoNT-A from serum samples of limited volume in most laboratory settings. Full article
(This article belongs to the Collection Rapid Detection of Bacterial Toxins)
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11 pages, 697 KiB  
Article
Detection of Shiga Toxins by Lateral Flow Assay
by Kathryn H. Ching, Xiaohua He, Larry H. Stanker, Alice V. Lin, Jeffery A. McGarvey and Robert Hnasko
Toxins 2015, 7(4), 1163-1173; https://doi.org/10.3390/toxins7041163 - 3 Apr 2015
Cited by 27 | Viewed by 8185
Abstract
Shiga toxin-producing Escherichia coli (STEC) produce shiga toxins (Stxs) that can cause human disease and death. The contamination of food products with STEC represents a food safety problem that necessitates rapid and effective detection strategies to mitigate risk. In this manuscript, we report [...] Read more.
Shiga toxin-producing Escherichia coli (STEC) produce shiga toxins (Stxs) that can cause human disease and death. The contamination of food products with STEC represents a food safety problem that necessitates rapid and effective detection strategies to mitigate risk. In this manuscript, we report the development of a colorimetric lateral flow assay (LFA) for the rapid detection of Stxs in <10 min using a pair of monoclonal antibodies that bind epitopes common to Stx1 and six Stx2 variants. This LFA provides a rapid and sensitive test for the detection of Stxs directly from STEC culture supernatants or at risk food samples with a 0.1 ng/mL limit of detection (LOD) for Stx2a. This Stx LFA is applicable for use in the rapid evaluation of Stx production from cultured E. coli strains or as a tool to augment current methods as part of food safety testing. Full article
(This article belongs to the Collection Shiga Toxins)
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15 pages, 496 KiB  
Article
A Monoclonal Antibody Based Capture ELISA for Botulinum Neurotoxin Serotype B: Toxin Detection in Food
by Larry H. Stanker, Miles C. Scotcher, Luisa Cheng, Kathryn Ching, Jeffery McGarvey, David Hodge and Robert Hnasko
Toxins 2013, 5(11), 2212-2226; https://doi.org/10.3390/toxins5112212 - 18 Nov 2013
Cited by 21 | Viewed by 9199
Abstract
Botulism is a serious foodborne neuroparalytic disease, caused by botulinum neurotoxin (BoNT), produced by the anaerobic bacterium Clostridium botulinum. Seven toxin serotypes (A–H) have been described. The majority of human cases of botulism are caused by serotypes A and B followed by [...] Read more.
Botulism is a serious foodborne neuroparalytic disease, caused by botulinum neurotoxin (BoNT), produced by the anaerobic bacterium Clostridium botulinum. Seven toxin serotypes (A–H) have been described. The majority of human cases of botulism are caused by serotypes A and B followed by E and F. We report here a group of serotype B specific monoclonal antibodies (mAbs) capable of binding toxin under physiological conditions. Thus, they serve as capture antibodies for a sandwich (capture) ELISA. The antibodies were generated using recombinant peptide fragments corresponding to the receptor-binding domain of the toxin heavy chain as immunogen. Their binding properties suggest that they bind a complex epitope with dissociation constants (KD’s) for individual antibodies ranging from 10 to 48 × 10−11 M. Assay performance for all possible combinations of capture-detector antibody pairs was evaluated and the antibody pair resulting in the lowest level of detection (L.O.D.), ~20 pg/mL was determined. Toxin was detected in spiked dairy samples with good recoveries at concentrations as low as 0.5 pg/mL and in ground beef samples at levels as low as 2 ng/g. Thus, the sandwich ELISA described here uses mAb for both the capture and detector antibodies (binding different epitopes on the toxin molecule) and readily detects toxin in those food samples tested. Full article
(This article belongs to the Special Issue Advances in Toxin Detection)
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