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Issue 20, 2014
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Development of an ELISA microarray assay for the sensitive and simultaneous detection of ten biodefense toxins

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Abstract

Plant and microbial toxins are considered bioterrorism threat agents because of their extreme toxicity and/or ease of availability. Additionally, some of these toxins are increasingly responsible for accidental food poisonings. The current study utilized an ELISA-based protein antibody microarray for the multiplexed detection of ten biothreat toxins, botulinum neurotoxins (BoNT) A, B, C, D, E, F, ricin, shiga toxins 1 and 2 (Stx), and staphylococcus enterotoxin B (SEB), in buffer and complex biological matrices. The multiplexed assay displayed a sensitivity of 1.3 pg mL−1 (BoNT/A, BoNT/B, SEB, Stx-1 and Stx-2), 3.3 pg mL−1 (BoNT/C, BoNT/E, BoNT/F) and 8.2 pg mL−1 (BoNT/D, ricin). All assays demonstrated high accuracy (75–120 percent recovery) and reproducibility (most coefficients of variation <20%). Quantification curves for the ten toxins were also evaluated in clinical samples (serum, plasma, nasal fluid, saliva, stool, and urine) and environmental samples (apple juice, milk and baby food) with overall minimal matrix effects. The multiplex assays were highly specific, with little cross-reactivity observed between the selected toxin antibodies. The results demonstrate a multiplex microarray that improves current immunoassay sensitivity for biological warfare agents in buffer, clinical, and environmental samples.

Graphical abstract: Development of an ELISA microarray assay for the sensitive and simultaneous detection of ten biodefense toxins

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Publication details

The article was received on 14 Jul 2014, accepted on 29 Jul 2014 and first published on 30 Jul 2014


Article type: Paper
DOI: 10.1039/C4AN01270D
Author version available: Download Author version (PDF)
Citation: Analyst, 2014,139, 5093-5102
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    Development of an ELISA microarray assay for the sensitive and simultaneous detection of ten biodefense toxins

    K. L. Jenko, Y. Zhang, Y. Kostenko, Y. Fan, C. Garcia-Rodriguez, J. Lou, J. D. Marks and S. M. Varnum, Analyst, 2014, 139, 5093
    DOI: 10.1039/C4AN01270D

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