Attomolar Detection of Botulinum Toxin Type A in Complex Biological Matrices

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2008 May 1

PMID 18446228

Citations 36

Authors

Karine Bagramyan

Jason R Barash

Stephen S Arnon

Markus Kalkum

Affiliations

Soon will be listed here.

Abstract

Background: A highly sensitive, rapid and cost efficient method that can detect active botulinum neurotoxin (BoNT) in complex biological samples such as foods or serum is desired in order to 1) counter the potential bioterrorist threat 2) enhance food safety 3) enable future pharmacokinetic studies in medical applications that utilize BoNTs.

Methodology/principal Findings: Here we describe a botulinum neurotoxin serotype A assay with a large immuno-sorbent surface area (BoNT/A ALISSA) that captures a low number of toxin molecules and measures their intrinsic metalloprotease activity with a fluorogenic substrate. In direct comparison with the "gold standard" mouse bioassay, the ALISSA is four to five orders of magnitudes more sensitive and considerably faster. Our method reaches attomolar sensitivities in serum, milk, carrot juice, and in the diluent fluid used in the mouse assay. ALISSA has high specificity for the targeted type A toxin when tested against alternative proteases including other BoNT serotypes and trypsin, and it detects the holotoxin as well as the multi-protein complex form of BoNT/A. The assay was optimized for temperature, substrate concentration, size and volume proportions of the immuno-sorbent matrix, enrichment and reaction times. Finally, a kinetic model is presented that is consistent with the observed improvement in sensitivity.

Conclusions/significance: The sensitivity, specificity, speed and simplicity of the BoNT ALISSA should make this method attractive for diagnostic, biodefense and pharmacological applications.

Citing Articles

An Advanced Sensing Approach to Biological Toxins with Localized Surface Plasmon Resonance Spectroscopy Based on Their Unique Protein Quaternary Structures.

Uzawa H, Kondo S, Nagatsuka T, Seto Y, Nishida Y Int J Mol Sci. 2025; 25(24.

PMID: 39769119 PMC: 11677682. DOI: 10.3390/ijms252413352.

International Proficiency Test Targeting a Large Panel of Botulinum Neurotoxin Sero- and Subtypes in Different Matrices.

Rasetti-Escargueil C, Popoff M, Kampa B, Worbs S, Marechal M, Guerin D Toxins (Basel). 2024; 16(11).

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Botulinum toxin as an ultrasensitive reporter for bacterial and SARS-CoV-2 nucleic acid diagnostics.

Song F, Shen Y, Wei Y, Yang C, Ge X, Wang A Biosens Bioelectron. 2021; 176:112953.

PMID: 33418182 PMC: 7836976. DOI: 10.1016/j.bios.2020.112953.

Toxemia in Human Naturally Acquired Botulism.

Rasetti-Escargueil C, Lemichez E, Popoff M Toxins (Basel). 2020; 12(11).

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Optimization of SNAP-25 and VAMP-2 Cleavage by Botulinum Neurotoxin Serotypes A-F Employing Taguchi Design-of-Experiments.

von Berg L, Stern D, Weisemann J, Rummel A, Dorner M, Dorner B Toxins (Basel). 2019; 11(10).

PMID: 31614566 PMC: 6832249. DOI: 10.3390/toxins11100588.

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