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Development of a separation platform comprising magnetic beads combined with the CRISPR/Cas12a system enabling ultrasensitive and rapid detection of miRNA-21

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Abstract

A separation platform has been developed mediated by a combination of magnetic beads and the CRISPR/Cas12a system to achieve ultrasensitive and rapid detection of miRNA-21 at a low level. In this system, with the assistance of an auxiliary probe, the target miRNA-21 can be specifically combined with three-stranded probes to initiate the SDR reaction. Abundant aptamer A3 was added to the solution that can activate the CRISPR/Cas12a system and initiate the trans-cleavage reaction to recover the fluorescence signal. Using magnetic beads to mediate the separation considerably greatly improves the signal conversion efficiency and detection sensitivity. At the 492 nm excitation wavelength, and 502–650 nm scan range, through analyzing the fluorescence peak intensity at 520 nm, the biosensor’s determination range and limit of detection is 8 fM-250 nM and 2.42 fM, respectively, and the RSD is 19.03–37.80. Compared with other biosensors, the biosensor developed exhibited superior performance and the signal recovered excellently in 1% human serum and the LOD is 12.12 fM. This method provides a novel highly sensitive scheme for detecting miRNA .

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (NSFC) (Grant No: 22164011, 21565015, 21663014), and the State Key Laboratory of Chemical Biosensing & Chemometrics.

Funding

National Natural Science Foundation of China,22164011,hongbo li

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Authors and Affiliations

  1. Key Lab of Fluorine and Silicon for Energy Materials and Chemistry of Ministry of Education, College of Chemistry and Chemical Engineering, Jiangxi Normal University, Nanchang, 330022, People’s Republic of China

    Xinyi Zhang, Hongbo Li, Weihua Zhao, Jun Xu & Suqin Wang

  2. State Key Laboratory for Chemo/Biosensing and Chemometrics, Hunan University, Changsha, 410082, People’s Republic of China

    Hongbo Li & Ruqin Yu

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  1. Xinyi Zhang
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  2. Hongbo Li
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  3. Weihua Zhao
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Correspondence to Hongbo Li.

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Zhang, X., Li, H., Zhao, W. et al. Development of a separation platform comprising magnetic beads combined with the CRISPR/Cas12a system enabling ultrasensitive and rapid detection of miRNA-21. Microchim Acta 190, 458 (2023). https://doi.org/10.1007/s00604-023-06038-w

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