DETECTION OF BOTULINUM NEUROTOXIN USING SYNTHETIC APTAMERS IN FLUORESCENCE POLARIZATION METHOD

Abstract

Aptamers are single strand oligonucleotides (DNA or RNA) that can detect any antigen in a sample matrix. Aptamers can be developed and optimized via Systematic Evolution of Ligand via Exponential Enrichment (SELEX) method. Previous studies have shown that aptamers have comparable specificity and affinity as antibodies used in conventional detection methods such as ELISA and lateral flow immunoassay (Bouvet, 2001). Using these aptamers, this project will attempt to develop a new detection method for botulinum neurotoxin (BoNT), which is the most toxic substance to human with LD50 as low as 2 ng/kg. Fluorescence Polarization (FP) method is based on polarization of light that can be deployed using aptamers to detect BoNT. The specific aims of this projects are as follow: 1. Develop two aptamers using CE-SELEX to detect BoNT A. 2. Using the two developed aptamers in conjunction with Fluorescence Polarization method to detect and quantify concentration of BoNT A in samples.