2013
DOI: 10.1021/jf3041963
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Detection of Botulinum Neurotoxin Serotypes A and B Using a Chemiluminescent versus Electrochemiluminescent Immunoassay in Food and Serum

Abstract: Botulinum neurotoxins (BoNT) are some of the most potent biological toxins. High-affinity monoclonal antibodies (mAbs) have been developed for the detection of BoNT serotypes A and B using a chemiluminescent capture ELISA. In an effort to improve toxin detection levels in complex matrices such as food and sera, we evaluated the performance of existing anti-toxin mAbs using a new electrochemiluminescence (ECL) immunoassay developed by Meso Scale Diagnostic instrument. In side-by-side comparisons, the limit of d… Show more

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Cited by 48 publications
(40 citation statements)
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“…The detection limit for BoNT/A was observed as 40 pg mL −1 (Fig. C), and our data were in a similar range to that published by others for BoNTs using ECL assays . In general, a lower sensitivity of the ECL assay was observed for all three BTAs in chocolate and raw milks compared with other tested milk products.…”
Section: Resultssupporting
confidence: 89%
See 1 more Smart Citation
“…The detection limit for BoNT/A was observed as 40 pg mL −1 (Fig. C), and our data were in a similar range to that published by others for BoNTs using ECL assays . In general, a lower sensitivity of the ECL assay was observed for all three BTAs in chocolate and raw milks compared with other tested milk products.…”
Section: Resultssupporting
confidence: 89%
“…Bacillus anthracis , staphylococcal enterotoxin B (SEB) and botulinum neurotoxins (BoNTs) are considered as potential bio threat agents (BTAs) that pose real threats to public health . Bacillus anthracis is the etiologic agent of anthrax, a disease primarily of herbivores and accidently, or intentionally, of humans .…”
Section: Introductionmentioning
confidence: 99%
“…For this purpose, methods that are based on the specific proteolytic activity of BoNTs have been commonly used to determine the serotype. In these methods, the detection signal is based on principles of fluorescence (Grate et al, 2009), chemiluminescence (Cadieux et al, 2005), electrochemiluminescence (Cheng and Stanker, 2013), surface plasmon resonance (Homola, 2003), or mass spectrometry (Kalb et al, 2010).…”
Section: Introductionmentioning
confidence: 99%
“…Milk matrices (non-fat, 2 %, whole milk) slightly increased the LOD to < 0.175 -0.314 ng, which was not statistically significant compared to assay buffer. Similar to previous work with other technologies such as ELISA and ECL [19], sample preparation of acidic juice (orange, apple, carrot) required neutralization of pH for the CANARY ® assay. Neutralized acidic juice samples were not centrifuged to remove particulate matter as used previously with other technologies but a matrix removal step was added using a magnetic bead separator before biosensors were added.…”
Section: Discussionmentioning
confidence: 96%
“…The protocol is similar to the one used for assay buffer and milk matrices. However, there are two key modifications: 1) Acidic juices were neutralized with 5 M Tris pH 8.0 (10% final volume) [19] before spiking and serial dilution with toxin. 2) After the 30 minute reaction to allow for binding of toxin:matrix with immunomagnetic capture beads; a magnetic separator was used to capture the complex, removal of the matrix, and replacement with an equal volume of assay buffer.…”
Section: Modified Canary ® Protocol For Acidic Juices: Carrot Applementioning
confidence: 99%