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A sandwich dipstick assay for ATP detection based on split aptamer fragments

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Abstract

Aptamer-based strip assay is an easy, highly efficient and low-cost detection method, which has been developed and easily applied to onsite detection. A new sensitive sandwich dipstick assay for adenosine triphosphate (ATP) detection was successfully developed based on specific recognition between split aptamer fragments and the target. In this method, the thiolated aptamer was first conjugated to the surface of gold nanoparticles (AuNPs), while the biotin-aptamer was immobilized on the surface of a nitrocellulose filter in the test line. In the presence of ATP, the thiol-aptamer/ATP/biotin-aptamer complexes were generated, which led to an obvious increase in optical signals at the test line. Under the optimal determination conditions, an excellent linear logarithmic response to the ATP concentration was obtained within the range of 0.5 μM to 5 mM. The limit of detection (LOD) of 0.5 μM was reached at a signal-to-noise ratio of 3. The dipstick assay showed a good average recovery of 96–108 % with the RSD of less than 20 % in urine samples. The proposed method exhibited high specificity against other nucleotides such as the uridine triphosphate (UTP), cytidine triphosphate (CTP), and guanosine triphosphate (GTP). The results indicated that the dipstick strip may be considered as an inexpensive screening tool for onsite ATP determination.

A simple split aptamer fragments based sandwich-type dipstick assay was developed for ATP detection

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Acknowledgments

All authors gratefully acknowledge the funding support from the Special Fund for Agro Scientific Research in the Public Interest (201203046, 201203023).

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    Authors and Affiliations

    1. Institute of Quality Standards and Testing Technology for Agro-products, Key Laboratory of Agro-product Quality and Safety, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

      Chao Zhu, Yan Zhao, Mengmeng Yan, Yafei Huang, Jiao Yan, Wenhui Bai & Ailiang Chen

    2. Key Laboratory of Agri-food Quality and Safety, Ministry of Agriculture, Beijing, 100081, China

      Chao Zhu, Yan Zhao, Mengmeng Yan, Yafei Huang, Jiao Yan, Wenhui Bai & Ailiang Chen

    3. College of Food Science and Technology, Hainan University, Haikou, Hainan, 570228, China

      Yafei Huang & Jiao Yan

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    1. Chao Zhu
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    Correspondence to Ailiang Chen.

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    Chao Zhu and Yan Zhao contributed equally to this work.

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    Zhu, C., Zhao, Y., Yan, M. et al. A sandwich dipstick assay for ATP detection based on split aptamer fragments. Anal Bioanal Chem 408, 4151–4158 (2016). https://doi.org/10.1007/s00216-016-9506-z

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