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A PCR-free voltammetric telomerase activity assay using a substrate primer on a gold electrode and DNA-triggered capture of gold nanoparticles

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Abstract

The paper describes a voltammetric method for the quantitation of the activity of telomerase extracted from cancer cells. A thiolated single-stranded telomerase substrate primer was firstly immobilized on a gold electrode. In the presence of a mixture of telomerase and deoxynucleotide triphosphates, the primer becomes elongated and contains repetitive nucleotide sequences (TTAGGG)n. After hybridization with blocker DNA, gold nanoparticles are added and captured by the elongated single-stranded DNA. This reduces the charge transfer resistance of the gold electrode. The telomerase activity is then quantified via differential pulse voltammetry, typically at 0.12 V (vs. SCE). The method is PCR-free, rapid, and convenient. It was applied to the detection of HeLa cells via the telomerase activity of lysed cells. The detection range was from 500 to 50,000 cells/mL and the detection limit was as low as 500 cells/mL.

A telomerase substrate (TS) primer is immobilized on a gold electrode as the sensing interface to detect the activity of telomerase extracted from cancer cells. Unmodified gold nanoparticles (AuNPs) are utilized which change the electrochemical responses.

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Acknowledgements

This work is supported by the National Natural Science Foundation of China (Grant no. 81771929) and the National Key Instrument Developing Project of China (Grant no. ZDYZ2013-1).

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Authors and Affiliations

  1. Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, 215163, People’s Republic of China

    Fanyu Meng, Wenfei Dong, Yuguo Tang & Peng Miao

  2. University of Science and Technology of China, Hefei, 230026, People’s Republic of China

    Fanyu Meng & Peng Miao

  3. Key Laboratory of Animal Physiology and Biochemistry, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, People’s Republic of China

    Yuanyuan Xu

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  1. Fanyu Meng
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  2. Yuanyuan Xu
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  4. Yuguo Tang
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  5. Peng Miao
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Correspondence to Peng Miao.

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Meng, F., Xu, Y., Dong, W. et al. A PCR-free voltammetric telomerase activity assay using a substrate primer on a gold electrode and DNA-triggered capture of gold nanoparticles. Microchim Acta 185, 398 (2018). https://doi.org/10.1007/s00604-018-2936-x

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  • DOI: https://doi.org/10.1007/s00604-018-2936-x

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