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Direct fluorescence anisotropy assay for cocaine using tetramethylrhodamine-labeled aptamer

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Abstract

Development of simple, sensitive, and rapid method for cocaine detection is important in medicine and drug abuse monitoring. Taking advantage of fluorescence anisotropy and aptamer, this study reports a direct fluorescence anisotropy (FA) assay for cocaine by employing an aptamer probe with tetramethylrhodamine (TMR) labeled on a specific position. The binding of cocaine and the aptamer causes a structure change of the TMR-labeled aptamer, leading to changes of the interaction between labeled TMR and adjacent G bases in aptamer sequence, so FA of TMR varies with increasing of cocaine. After screening different labeling positions of the aptamer, including thymine (T) bases and terminals of the aptamer, we obtained a favorable aptamer probe with TMR labeled on the 25th base T in the sequence, which exhibited sensitive and significant FA-decreasing responses upon cocaine. Under optimized assay conditions, this TMR-labeled aptamer allowed for direct FA detection of cocaine as low as 5 μM. The maximum FA change reached about 0.086. This FA method also enabled the detection of cocaine spiked in diluted serum and urine samples, showing potential for applications.

The binding of cocaine to the TMR-labeled aptamer causes conformation change and alteration of the intramolecular interaction between TMR and bases of aptamer, leading to variance of fluorescence anisotropy (FA) of TMR, so direct FA analyis of cocaine is achieved

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Acknowledgements

We acknowledged the support from National Natural Science Foundation of China (Grant No. 21575153, 21435008), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB14030200), and the Key Research Program of the Chinese Academy of Sciences (KFZD-SW-203).

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Authors and Affiliations

  1. State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, 100085, China

    Yingxiong Liu & Qiang Zhao

  2. Institute of Environmental Science, College of Chemistry and Chemical Engineering, Shanxi University, Taiyuan, Shanxi, 030006, China

    Yingxiong Liu

  3. University of Chinese Academy of Sciences, Beijing, 100049, China

    Qiang Zhao

Authors
  1. Yingxiong Liu
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  2. Qiang Zhao
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Correspondence to Qiang Zhao.

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Liu, Y., Zhao, Q. Direct fluorescence anisotropy assay for cocaine using tetramethylrhodamine-labeled aptamer. Anal Bioanal Chem 409, 3993–4000 (2017). https://doi.org/10.1007/s00216-017-0349-z

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  • DOI: https://doi.org/10.1007/s00216-017-0349-z

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