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Label-free electrochemiluminescence biosensor for ultrasensitive detection of telomerase activity in HeLa cells based on extension reaction and intercalation of Ru(phen)3 2+

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Abstract

Telomerase is one of the most common markers of human malignant tumors, such as uterine, stomach, esophageal, breast, colorectal, laryngeal squamous cell, thyroid, bladder, and so on. It is necessary to develop some sensitive but convenient detection methods for telomerase activity determination. In this study, a label-free and ultrasensitive electrochemiluminescence (ECL) biosensor has been fabricated to detect the activity of telomerase extracted from HeLa cells. Thiolated telomerase substrate (TS) primer was immobilized on the gold electrode surface through gold-sulfur (Au-S) interaction and then elongated by telomerase specifically. Then, it was hybridized with complementary DNA to form double-stranded DNA (dsDNA) fragments on the electrode surface, and Ru(phen)3 2+ has been intercalated into the dsDNA grooves to act as the ECL probe. The enhanced ECL intensity has a linear relationship with the number of HeLa cells in the range of 5∼5000 and with a detection limit of 2 HeLa cells. The proposed ECL biosensor has high specificity to telomerase in the presence of common interferents. The relative standard deviations (RSDs) were <5 % at 100 HeLa cells. The proposed method provides a convenient approach for telomerase-related cancer screening or diagnosis.

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Acknowledgments

This project was partly financially supported by the National Sciences Foundation of China (21275031, 21575027, 21305141), Program for New Century Excellent Talents in University (NCET-12-0619), Natural Sciences Funding of Fujian Province (2014J06005), and Program for Changjiang Scholars and Innovative Research Team in University (No. IRT15R11).

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Authors and Affiliations

  1. Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Fuzhou University, Fuzhou, Fujian, 350116, China

    Yue Lin, Linlin Yang, Guiyin Yue, Bin Qiu, Longhua Guo, Zhenyu Lin & Guonan Chen

  2. State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian, 350002, China

    Lifen Chen

  3. Department of Chemistry, Fuzhou University, Fuzhou, Fujian, 350116, China

    Zhenyu Lin

Authors
  1. Yue Lin
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  2. Linlin Yang
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  3. Guiyin Yue
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  4. Lifen Chen
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  5. Bin Qiu
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  6. Longhua Guo
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  7. Zhenyu Lin
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  8. Guonan Chen
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Correspondence to Zhenyu Lin.

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Additional information

Published in the topical collection Analytical Electrochemiluminescence with guest editors Hua Cui, Francesco Paolucci, Neso Sojic, and Guobao Xu.

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Lin, Y., Yang, L., Yue, G. et al. Label-free electrochemiluminescence biosensor for ultrasensitive detection of telomerase activity in HeLa cells based on extension reaction and intercalation of Ru(phen)3 2+ . Anal Bioanal Chem 408, 7105–7111 (2016). https://doi.org/10.1007/s00216-016-9561-5

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  • DOI: https://doi.org/10.1007/s00216-016-9561-5

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