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(-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.
authors.affiliation: Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA 99354, USA. susan.varnum@pnnl.gov.
authors.forename: Kathryn L
authors.initials: KL
authors.lastname: Jenko
authors.forename: Yanfeng
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yulia
authors.initials: Y
authors.lastname: Kostenko
authors.forename: Yongfeng
authors.initials: Y
authors.lastname: Fan
authors.forename: Consuelo
authors.initials: C
authors.lastname: Garcia-Rodriguez
authors.forename: Jianlong
authors.initials: J
authors.lastname: Lou
authors.forename: James D
authors.initials: JD
authors.lastname: Marks
authors.forename: Susan M
authors.initials: SM
authors.lastname: Varnum
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D001905:Botulinum Toxins / Q000032:analysis; D004768:Enterotoxins / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D008892:Milk / Q000737:chemistry; D040081:Protein Array Analysis; D012276:Ricin / Q000032:analysis; D022603:Shiga Toxins / Q000032:analysis; D014116:Toxicology / Q000379:methods*; D014118:Toxins, Biological / Q000032:analysis* / Q000097:blood / Q000652:urine
chemical_list: D000906:Antibodies; D004768:Enterotoxins; D022603:Shiga Toxins; D014118:Toxins, Biological; C031818:enterotoxin B, staphylococcal; D012276:Ricin; D001905:Botulinum Toxins
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