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Amplified Analysis of DNA or Proteins by TdT-generated DNAzyme

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Abstract

Sensitive and specific detection of nucleic acids or proteins, which act as biomarkers, is of great importance in disease diagnosis. By combing the concept and operation of an endonuclease-assisted target-responsive amplification method and peroxidase-mimic DNAzyme generated by terminal deoxynucleotidyl transferase (TdT), a novel and facile colorimetric biosensor was developed for DNA and protein. Target DNA and thrombin were chosen as representative biomolecules. The production of cleavage fragments can only be triggered by specific target binding and the following nicking process, which do not occur spontaneously. In the signal collection part, numerous guanine-rich DNA were produced through the prolongation of cleavage fragments by TdT and formed highly effective DNAzyme with hemin. In this novel amplification method, we succeeded in realizing sensitive and specific detection of target DNA and thrombin. Under optimal conditions, target DNA can be detected as low as 1 pM, and thrombin with a detection limit of 100 pM. The method also proves the potential versatility and feasibility of TdT-generated DNAzyme in various bio-analyses.

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Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (21705163, 21601204), Natural Science Foundation of Hunan Province (2019JJ50737), the Research Project of the National University of Defense Technology (ZK18-03-39) and the Open Subject of State Key Laboratory of Chemo/Biosensing and Chemometrics (2017011).

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Authors and Affiliations

  1. College of Liberal Arts and Sciences, National University of Defense Technology, Changsha, Hunan, 410073, China

    Zhuoliang Liu, Tianxiong Liu, Cheng-An Tao, Xianzhe Chen, Jian Huang, Fang Wang & Jianfang Wang

  2. State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, Hunan, 410082, China

    Zhuoliang Liu

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  1. Zhuoliang Liu
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  2. Tianxiong Liu
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  3. Cheng-An Tao
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  4. Xianzhe Chen
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  5. Jian Huang
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  6. Fang Wang
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  7. Jianfang Wang
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Correspondence to Jianfang Wang.

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Liu, Z., Liu, T., Tao, CA. et al. Amplified Analysis of DNA or Proteins by TdT-generated DNAzyme. ANAL. SCI. 36, 835–840 (2020). https://doi.org/10.2116/analsci.19P387

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  • DOI: https://doi.org/10.2116/analsci.19P387

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