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Journal of Analytical Methods in Chemistry
Volume 2016, Article ID 9241860, 8 pages
http://dx.doi.org/10.1155/2016/9241860
Research Article

Preparation of a Specific ssDNA Aptamer for Brevetoxin-2 Using SELEX

1Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China
2Emergency Department, The Eastern Division, The First Hospital of Jilin University, Changchun 130062, China
3Fuqing Entry-Exit Inspection and Quarantine Bureau, Port District, Qingrong Road, Fuqing, Fujian 350300, China

Received 10 August 2016; Revised 20 October 2016; Accepted 30 October 2016

Academic Editor: Beate Strehlitz

Copyright © 2016 Rui-Yun Tian et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The existing assays for detecting brevetoxin (BTX) depend on expensive equipment with a professional operator or on an antibody with limited stability, which requires complex processes, a high cost, and a considerable amount of time. The development of an alternative detection probe is another promising research direction. This paper reports the use of aptamers binding to BTX-2 in an analytical assay using the systematic evolution of ligands by exponential enrichment (SELEX). After 12 rounds of selection, the secondary structures of 25 sequences were predicted. Compared to other aptamers, Bap5 has relatively high affinity with the lowest dissociation constant of 4.83 μM, and IC50 is 73.81 ng mL−1. A good linear regression formula of with a coefficient correlation of = 0.9798 was obtained using a biotin-avidin ELISA. Moreover, there is no cross-reaction with the detected marine toxins, except for BTX-2. Thus, Bap5 has potential to detect BTX-2 in shellfish in the future as a substitute for the recognition probe.