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United States Department of Agriculture

Agricultural Research Service

Research Project: Immunodiagnostics to Detect Prions and Other Important Animal Pathogens

Location: Produce Safety and Microbiology Research

Title: Rapid microfluidic assay for the detection of botulinum neurotoxin in animal sera

Author
item Babrak, Lmar
item Lin, Alice
item Stanker, Larry
item Mcgarvey, Jeffery - Jeff
item Hnasko, Robert

Submitted to: Toxins
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/25/2015
Publication Date: 1/4/2016
Citation: Babrak, L.M., Lin, A.V., Stanker, L.H., McGarvey, J.A., Hnasko, R.M. 2016. Rapid microfluidic assay for the detection of botulinum neurotoxin in animal sera. Toxins. 8(1):13.

Interpretive Summary: Botulism caused by botulinum neurotoxins (BoNTs) represents a threat to public health and safety. We have developed a rapid high-throughput microfluid BoNT immunoassay that detects BoNT from very small amount of animal sera in 75 minutes with minimal biohazardous waste. This improves existing assays by reducing required sample volume, time, and waste disposal.

Technical Abstract: The potent botulinum neurotoxins (BoNTs) represent a threat to public health and safety. Botulism is a disease caused by BoNT intoxication that results in muscle paralysis that can be fatal. Sensitive assays capable of detecting BoNTs from different substrates and settings are essential to limit foodborne contamination and morbidity. In this report, we describe a rapid 96-well microfluidic double sandwich immunoassay for the sensitive detection of BoNT-A from animal sera. This BoNT microfluidic assay requires only 5 µL of serum, provides results in 75 minutes using a standard fluorescence microplate reader and generates minimal hazardous waste. The assay has a < 30 pg ml-1 limit of detection (LOD) of BoNT-A from spiked human serum. This sensitive microfluidic BoNT-A assay offers a fast and simplified workflow suitable for the detection of BoNT-A from serum samples of limited volume in most laboratory settings.

Last Modified: 09/24/2017
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