title: In vitro detection and quantification of botulinum neurotoxin type e activity in avian blood.
abstract: Botulinum neurotoxin serotype E (BoNT/E) outbreaks in the Great Lakes region cause large annual avian mortality events, with an estimated 17,000 bird deaths reported in 2007 alone. During an outbreak investigation, blood collected from bird carcasses is tested for the presence of BoNT/E using the mouse lethality assay. While sensitive, this method is labor-intensive and low throughput and can take up to 7 days to complete. We developed a rapid and sensitive in vitro assay, the BoTest Matrix E assay, that combines immunoprecipitation with high-affinity endopeptidase activity detection by Förster resonance energy transfer (FRET) to rapidly quantify BoNT/E activity in avian blood with detection limits comparable to those of the mouse lethality assay. On the basis of the analysis of archived blood samples (n = 87) collected from bird carcasses during avian mortality investigations, BoTest Matrix E detected picomolar quantities of BoNT/E following a 2-h incubation and femtomolar quantities of BoNT/E following extended incubation (24 h) with 100% diagnostic specificity and 91% diagnostic sensitivity.
authors: Department of Forest and Wildlife Ecology, University of Wisconsin, 204 Russell Labs, 1630 Linden Drive, Madison, Wisconsin 53706, USA.
authors: Timothy M
authors: TM
authors: Piazza
authors: David S
authors: DS
authors: Blehert
authors: F Mark
authors: FM
authors: Dunning
authors: Brenda M
authors: BM
authors: Berlowski-Zier
authors: Füsûn N
authors: FN
authors: Zeytin
authors: Michael D
authors: MD
authors: Samuel
authors: Ward C
authors: WC
authors: Tucker
mesh_terms: D000818:Animals; D001715:Bird Diseases / Q000175:diagnosis*; D001717:Birds; D001774:Blood Chemical Analysis / Q000379:methods*; D001905:Botulinum Toxins / Q000097:blood*; D001906:Botulism / Q000175:diagnosis / Q000662:veterinary*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D006114:Great Lakes Region; D047468:Immunoprecipitation / Q000379:methods; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D001905:Botulinum Toxins; C096326:botulinum toxin type E