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Construction of Dual-Target Recognition-Based Specific MicroRNA Detection Method for Acute Pancreatitis Analysis

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Abstract

MicroRNAs (miRNAs) play crucial roles in regulating various biological processes and are considered promising biomarkers for clinical diagnosis and therapy of acute pancreatitis. Herein, we present a duplex-specific nuclease (DSN enzyme) and DNAzyme-assisted fluorescent miRNA detections assay that can provide improved detection specificity due to a design of dual-target recognition and a comparable sensitivity. The dual-target recognitions are composed of (i) miRNA unfold hairpin structure toehold to form DNA-RNA duplex, among which the DNA section will be digested by DSN enzyme, releasing miRNA to participant in a next recycle. (ii) After DNAzyme-based nicking site formation in loop section of molecular beacon (MB), miRNA can bind with the loop section of MB and gradually unfold MB probe, generating fluorescence signals. With this general principle, distinct discrimination capability towards even one base pair mismatch of homogenous miRNA is obtained, showing a promising prospect in clinical diagnosis and therapy of acute pancreatitis.

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All data generated and analyzed during this study are included in this article.

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Acknowledgements

The authors thank the financial and equipment support from The Third Affiliated Hospital of Chongqing Medical University.

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Authors and Affiliations

  1. Emergency Department of The Third Affiliated Hospital of Chongqing Medical University, 1 Huixing Shuanghu Branch Road, Yubei District, Chongqing, 401120, China

    Chunhong Sun, Yuan Rong, Zeping Yang, Dong She & Mingwei Gong

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  1. Chunhong Sun
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  2. Yuan Rong
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  3. Zeping Yang
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  4. Dong She
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  5. Mingwei Gong
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Contributions

S.C. and G.M. designed the strategy, completed the preparation of the research, and wrote the manuscript; R.Y., Y.Z., S.D., and G.M. assisted data analysis.

Corresponding author

Correspondence to Mingwei Gong.

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Sun, C., Rong, Y., Yang, Z. et al. Construction of Dual-Target Recognition-Based Specific MicroRNA Detection Method for Acute Pancreatitis Analysis. Appl Biochem Biotechnol 194, 3136–3144 (2022). https://doi.org/10.1007/s12010-022-03907-7

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  • DOI: https://doi.org/10.1007/s12010-022-03907-7

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