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Int. J. Mol. Sci. 2018, 19(1), 85; https://doi.org/10.3390/ijms19010085

In Vitro Selection of a Single-Stranded DNA Molecular Recognition Element against the Pesticide Fipronil and Sensitive Detection in River Water

1
Department of Pharmaceutical Sciences, Nesbitt School of Pharmacy, Wilkes University, 84 W. South Street, Wilkes-Barre, PA 18766, USA
2
Independent Researcher, Pittsburgh, PA 15234, USA
*
Author to whom correspondence should be addressed.
Received: 31 October 2017 / Revised: 12 December 2017 / Accepted: 25 December 2017 / Published: 28 December 2017
(This article belongs to the Special Issue Aptamers)
View Full-Text   |   Download PDF [9769 KB, uploaded 28 December 2017]   |  

Abstract

Fipronil is a commonly used insecticide that has been shown to have environmental and human health risks. The current standard methods of detection for fipronil and its metabolites, such as GC-MS, are time consuming and labor intensive. In this study, a variant of systematic evolution of ligands by exponential enrichment (SELEX), was utilized to identify the first single-stranded DNA (ssDNA) molecular recognition element (MRE) that binds to fipronil with high affinity (Kd = 48 ± 8 nM). The selected MRE displayed low cross binding activity on various environmentally relevant, structurally unrelated herbicides and pesticides, in addition to broad-spectrum binding activity on major metabolites of fipronil and a structurally similar pesticide in prepared river samples. Additionally, a proof-of-principle fluorescent detection assay was developed by using the selected ssDNA MRE as a signal-reporting element, with a limit of detection of 105 nM in a prepared river water sample. View Full-Text
Keywords: SELEX; in vitro selection; aptamer; molecular recognition element (MRE); fipronil; pesticide; insecticide; ELISA; Decoy-SELEX SELEX; in vitro selection; aptamer; molecular recognition element (MRE); fipronil; pesticide; insecticide; ELISA; Decoy-SELEX
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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. (CC BY 4.0).

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Hong, K.L.; Sooter, L.J. In Vitro Selection of a Single-Stranded DNA Molecular Recognition Element against the Pesticide Fipronil and Sensitive Detection in River Water. Int. J. Mol. Sci. 2018, 19, 85.

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