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

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 (BoNT/A, BoNT/B, SEB, Stx-1 and Stx-2), 3.3 pg/mL (BoNT/C, BoNT/E, BoNT/F) and 8.2 pg/mL (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 crossreactivity 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.

Authors:
; ; ; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1167301
Report Number(s):
PNNL-SA-105212
400412000
DOE Contract Number:
AC05-76RL01830
Resource Type:
Journal Article
Resource Relation:
Journal Name: Analyst, 139(20):5093-5102
Country of Publication:
United States
Language:
English

Citation Formats

Jenko, Kathryn, Zhang, Yanfeng, Kostenko, Yulia, Fan, Yongfeng, Garcia-Rodriguez, Consuelo, Lou, Jianlong, Marks, James D., and Varnum, Susan M. Development of an ELISA microarray assay for the sensitive and simultaneous detection of ten biodefense toxins.. United States: N. p., 2014. Web. doi:10.1039/c4an01270d.
Jenko, Kathryn, Zhang, Yanfeng, Kostenko, Yulia, Fan, Yongfeng, Garcia-Rodriguez, Consuelo, Lou, Jianlong, Marks, James D., & Varnum, Susan M. Development of an ELISA microarray assay for the sensitive and simultaneous detection of ten biodefense toxins.. United States. doi:10.1039/c4an01270d.
Jenko, Kathryn, Zhang, Yanfeng, Kostenko, Yulia, Fan, Yongfeng, Garcia-Rodriguez, Consuelo, Lou, Jianlong, Marks, James D., and Varnum, Susan M. 2014. "Development of an ELISA microarray assay for the sensitive and simultaneous detection of ten biodefense toxins.". United States. doi:10.1039/c4an01270d.
@article{osti_1167301,
title = {Development of an ELISA microarray assay for the sensitive and simultaneous detection of ten biodefense toxins.},
author = {Jenko, Kathryn and Zhang, Yanfeng and Kostenko, Yulia and Fan, Yongfeng and Garcia-Rodriguez, Consuelo and Lou, Jianlong and Marks, James D. and Varnum, Susan M.},
abstractNote = {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 (BoNT/A, BoNT/B, SEB, Stx-1 and Stx-2), 3.3 pg/mL (BoNT/C, BoNT/E, BoNT/F) and 8.2 pg/mL (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 crossreactivity 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.},
doi = {10.1039/c4an01270d},
journal = {Analyst, 139(20):5093-5102},
number = ,
volume = ,
place = {United States},
year = 2014,
month =
}
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