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T790M mutation upconversion fluorescence biosensor via mild ATRP strategy and site-specific DNA cleavage of restriction endonuclease

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Abstract

A unique combination of a specific nucleic acid restriction endonuclease (REase) and atom transfer radical polymerization (ATRP) signal amplification strategy was employed for the detection of T790M mutations prevalent in the adjuvant diagnosis of lung cancer. REase selectively recognizes and cleaves T790M mutation sites on double-stranded DNA formed by hybridization of a capture sequence and a target sequence. At the same time, the ATRP strategy resulted in the massive aggregation of upconverted nanoparticles (UCNPs), which significantly improved the sensitivity of the biosensor. In addition, the UCNPs have excellent optical properties and can eliminate the interference of autofluorescence in the samples, thus further improving the detection sensitivity. The proposed upconversion fluorescent biosensor is characterized by high specificity, high sensitivity, mild reaction conditions, fast response time, and a detection limit as low as 0.14 fM. The performance of the proposed biosensor is comparable to that of clinical PCR methods when applied to clinical samples. This work presents a new perspective for assisted diagnosis in the pre-intervention stage of tumor diagnostics in the early stage of precision oncology treatments.

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Acknowledgements

This work was supported by the project of tackling of key scientific and technical problems in Henan Province (232102321119, 202102310149).

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Authors and Affiliations

  1. Pharmacy College, Henan University of Chinese Medicine, Zhengzhou, 450046, People’s Republic of China

    Yanju Liu, Liang Guo, Mengyuan Hou, Haiyang Gao, Yuanmeng Ke, Huaixia Yang & Fuchun Si

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Contributions

Conceptualization, Yanju Liu and Huaixia Yang; methodology, Yanju Liu and Liang Guo; validation, Liang Guo, and Mengyuan Hou; formal analysis, Hiayang Gao.; investigation, Yanju Liu, Huaixia Yang and Fuchun Si; resources, Huaixia Yang and Fuchun Si; writing-original draft preparation, Yanju Liu and Mengyuan Hou; writing-review and editing, Liang Guo and Yuanmeng Ke; supervision, Huaixia Yang; funding acquisition, Huaixia Yang and Yanju Liu. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Huaixia Yang or Fuchun Si.

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Liu, Y., Guo, L., Hou, M. et al. T790M mutation upconversion fluorescence biosensor via mild ATRP strategy and site-specific DNA cleavage of restriction endonuclease. Microchim Acta 191, 148 (2024). https://doi.org/10.1007/s00604-024-06229-z

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