Split aptamer-based sandwich fluorescence resonance energy transfer assay for 19-nortestosterone

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• 作者：Wenhui Bai ; Chao Zhu ; Jinchuan Liu ; Mengmeng Yan ; Shuming Yang…
• 刊名：Microchimica Acta
• 出版年：2016
• 出版时间：September 2016
• 年：2016
• 卷：183
• 期：9
• 页码：2533-2538
• 全文大小：533 KB
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Inorganic Chemistry
  Physical Chemistry
  Characterization and Evaluation Materials
  Monitoring, Environmental Analysis and Environmental Ecotoxicology
  
• 出版者：Springer Wien
• ISSN：1436-5073
• 卷排序：183

文摘

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.