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Sensitive naked-eye detection of telomerase activity based on exponential amplification reaction and lateral flow assay

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Abstract

Telomerase is a promising diagnostic and prognostic biomarker for cancers. Sensitive, simple, and reliable telomerase activity detection is vital for cancer diagnosis. Herein, we developed an ultrasensitive visualized assay for telomerase activity that combined the exponential amplification reaction (EXPAR) and lateral flow assay for easy and quick signal readout, which we termed as a lateral flow readout-EXPAR (LFR-EXPAR) assay. In the LFR-EXPAR assay, telomerase elongation products initiate the exponential amplification reaction, the generated trigger hybridizes with the reporter to form the recognition site of the nicking enzyme, and the nicking enzyme cuts the reporter strand. The degradation of the reporter can be detected with a universal lateral flow dipstick and read out with the naked eye. After conducting a series of proof-of-concept investigations, the LFR-EXPAR assay was found to achieve a sensitivity comparable to that of a TRAP (telomere repeat amplification protocol) assay. The LFR-EXPAR assay can be used to realize ultrasensitive and point-of-care detection of telomerase without requiring specialized instruments, holding great promise for early cancer diagnosis.

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Acknowledgements

We thank Yan Zhang (College of Chemistry, Shandong Normal University, Jinan 250014, China) and Qinfeng Xu (School of Food and Biological Engineering, Shaanxi University of Science and Technology, Xi’an 710021, China) for insightful discussion and suggestions.

Funding

This work was supported by the National Natural Science Foundation of China (NSFC) (grant number 32000866), the Major Research Program of the National Natural Science Foundation of China (grant number 91849209), and the Natural Science Foundation of Shandong Province (grant number ZR201910210189).

All animal experiments were performed in compliance with relevant laws or guidelines. All animal experiments followed the institutional guidelines of Qingdao University. The Ethics Committee of Qingdao University approved our studies. Informed consent was obtained from all participants included in the study.

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Authors and Affiliations

  1. Institute for Translational Medicine, The Affiliated Hospital of Qingdao University, College of Medicine, Qingdao University, 1 Ningde Road, Qingdao, 266073, China

    Xue-Ru Cheng, Fei Wang, Cui-yun Liu, Jing Li, Chan Shan, Kun Wang, Yin Wang, Pei-Feng Li & Xin-Min Li

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  1. Xue-Ru Cheng
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  2. Fei Wang
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  3. Cui-yun Liu
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  4. Jing Li
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  6. Kun Wang
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  7. Yin Wang
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  8. Pei-Feng Li
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  9. Xin-Min Li
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Contributions

Xue-Ru Cheng: methodology, investigation, validation, formal analysis, writing—original draft, and writing—review and editing. Cui-yun Liu: formal analysis, methodology, writing—review and editing. Fei Wang: formal analysis, methodology, writing—review and editing. Jing Li: formal analysis, methodology, writing—review and editing. Chan Shan: formal analysis, methodology, writing—review and editing. Kun Wang: formal analysis, methodology, writing—review and editing. Yin Wang: conceptualization, data curation, formal analysis, writing—review and editing. Pei-Feng Li: conceptualization, data curation, funding acquisition, formal analysis, writing—review and editing, supervision. Xin-min Li: conceptualization, methodology, funding acquisition, writing—original draft, writing—review and editing, project administration, supervision.

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Correspondence to Yin Wang, Pei-Feng Li or Xin-Min Li.

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Cheng, XR., Wang, F., Liu, Cy. et al. Sensitive naked-eye detection of telomerase activity based on exponential amplification reaction and lateral flow assay. Anal Bioanal Chem 414, 6139–6147 (2022). https://doi.org/10.1007/s00216-022-04179-0

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  • DOI: https://doi.org/10.1007/s00216-022-04179-0

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