Single-stranded DNA (ssDNA) aptamers against four organophosphorus pesticides (phorate, profenofos, isocarbophos and omethoate) were simultaneously isolated from an immobilized random ssDNA library by systematic evolution of ligands by exponential enrichment (SELEX) technique. After 12 rounds of in vitro selection, five ssDNA aptamer candidates were selected and their binding affinities were identified by a novel method using a molecular beacon. Two of the five ssDNA sequences, SS2-55 and SS4-54, demonstrated higher affinities and specificities to the four organophosphorus pesticides. They were defined as broad-spectrum aptamers binding to four different targets and their simulated secondary structures showed highly distinct features with typical stem and loop structures. The dissociation constant of SS2-55 and SS4-54 binding to the four organophosphorus pesticides ranged from 0.8 to 2.5 μM. These aptamers offered application potential in the analysis and/or neutralization of the residues of the four organophosphorus pesticides.
DNA aptamer Food safety monitoring and analysis Molecular beacon Organophosphorus pesticide SELEX
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This work was supported by research grants received from the National Natural Science Foundation of China (30871658), the “948” Projects of Chinese Agriculture Ministry (2011-Z46 and 2011-G5-7), the Independent Innovation Foundation of Jiangsu Province in China [CX (10)236], the Jingsu Planed Projects for Postdoctoral Research Funds (0802023B) and Suzhou Key Technology Research and Development Program (SN201129).
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