Real-time assay for the detection of botulinum toxin

摘要

A real-time portable and rapid detection assay to identify the presence of biologically active toxins such as botulinum toxins. The proteolytic activity of BoNT/A is measured using a peptide cleavage assay, where a fluorescent substrate is cleaved by BoNT/A, resulting in increased fluorescence. This fluorescence can be monitored in real-time using a fluorescence detection instrument, such as a real-time PCR system that has been modified to implement a detection algorithm specific to the identification of the target toxin.

主权项

1. A method for detecting the presence of biologically active botulinum toxin, comprising the steps of:
providing a peptide substrate comprising SEQ. ID NO. 1 labeled with a fluorophore and a quencher and having a cleavage site responsive to biologically active botulinum toxin positioned between said fluorophone and said quencher; adding a test sample to said peptide substrate so that any biologically active botulinum toxin contained in said test sample will cleave said peptide substrate, thereby allowing said peptide substrate to fluoresce; incubating said peptide substrate; detecting the amount of fluorescence emitted from said sample and outputting a corresponding fluorescence reading; determining whether any biologically active botulinum toxin is present in said sample based on said fluorescence reading.