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Appl Environ Microbiol. 2011 Nov;77(21):7815-22. doi: 10.1128/AEM.06165-11. Epub 2011 Sep 9.

In vitro detection and quantification of botulinum neurotoxin type e activity in avian blood.

Author information

1
Department of Forest and Wildlife Ecology, University of Wisconsin, 204 Russell Labs, 1630 Linden Drive, Madison, Wisconsin 53706, USA.

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.

PMID:
21908624
PMCID:
PMC3209138
DOI:
10.1128/AEM.06165-11
[Indexed for MEDLINE]
Free PMC Article

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