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.
Highlights
Fipronil has been a widely used phenlpyrazole insecticide in the United States since it was first introduced in the late 1990s [1]
Round 12 sequences were subjected to Mfold analysis for secondary structures with associated Gibbs free energy values (∆G)
It is important to note that this study showed the identification of a fipronil binding element without the use of a hapten or animals
Summary
Fipronil has been a widely used phenlpyrazole insecticide in the United States since it was first introduced in the late 1990s [1]. Fipronil is often used as an alternative to organophosphate pesticides in many settings, including residential, commercial, and agricultural, due to its selectivity on insect GABA receptors [3]. Fipronil has become a widespread environmental contaminant because of its wide applications. In the United States from 2002 to 2011, fipronil was detected in urban streams up to 63% of the time, and 15 to 20% of the time in agricultural and mixed land [3]. Measured concentrations of fipronil exceed the aquatic-life benchmark in 70% of urban streams and more than 20% of agricultural and mixed land streams in this timeframe [3]. Worldwide exposure of humans and ecosystems to fipronil has been clearly observed
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