1.Key Laboratory of Agro-product Quality and Safety, Institute of Quality Standards and Testing Technology for Agro-products,Chinese Academy of Agricultural Sciences,Beijing,China;2.Key Laboratory of Agri-food Quality and Safety, Ministry of Agriculture,Beijing,China
Abstract:
In order to develop an aptamer based fluorescence resonance energy transfer (FRET) assay for 19-nortestosterone, a 76-mer 17β-estradiol aptamer was split into two pieces (referred to as P1 and P2, respectively). P1 was labeled with a quencher (BHQ), and P2 with a fluorophore (6FAM). The two aptamer pieces were employed to detect NT via FRET quenching in a homogeneous solution. This method has a low detection limit (5 μM) within a wide dynamic range (5 to 1000 μM). The approach was used to analyze spiked urine samples, and the results showed that the average recovery of three samples containing different NT concentrations ranged from 58 to 118 % with a relative standard deviation (RSD) of less than 1 %. In our perception, the method has a wide scope for future applications to other analytes by using dually labeled split aptamers.
Graphical abstract A split aptamer-based fluorescence resonance energy transfer assay for 19-nortestosterone was developed with a wide dynamic range of 5 to 1000 μM and low detection limit (5 μM). The average recovery from spiked urine samples ranged from 58 to 118 %, with a relative standard deviation (RSD) of less than 1 %.
