In vitro detection and quantification of botulinum neurotoxin type E activity in avian blood
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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.
Study Area
Publication type | Article |
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Publication Subtype | Journal Article |
Title | In vitro detection and quantification of botulinum neurotoxin type E activity in avian blood |
Series title | Applied and Environmental Microbiology |
DOI | 10.1128/AEM.06165-11 |
Volume | 77 |
Issue | 21 |
Year Published | 2011 |
Language | English |
Publisher | American Society for Microbiology |
Contributing office(s) | National Wildlife Health Center |
Description | 8 p. |
First page | 7815 |
Last page | 7822 |
Country | Canada, United States |
State | Illinois, Indiana, Michigan, Minnesota, New York, Ohio, Ontario, Pennsylvania, Quebec, Wisconsin |
Other Geospatial | Great Lakes |