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Ultra-sensitive electrochemiluminescent biosensor for miRNA based on CRISPR/Cas13a trans-cleavage-triggered hybridization chain reaction and magnetic-assisted enrichment

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Abstract

The great selectivity and trans-cleavage activity of clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a had been coupled with high amplification efficiency of hybridization chain reaction (HCR) and magnetic-assisted enrichment, high sensitivity of electrochemiluminescence (ECL) detection to develop an ultra-sensitive biosensor for microRNA-21 (miRNA-21). The CRISPR/Cas13a was used to recognize target RNA with high specificity and performed the trans-cleavage activity. An initiation strand was generated to bind to the probe on the surface of nanomagnetic beads and then trigged HCR to produce long double-strand DNAs (dsDNAs) to realize signal amplification. Ru(phen)32+ can be inserted in the groove of the dsDNAs and acts as the ECL indicator, which can be separated through magnetic enrichment and allowed the platform to reduce the signal background. Under the optimized conditions, there is a good linear correlation between the ECL intensity and the logarithm of miRNA-21 concentration in the range 1 fM–10 nM; the limit of detection (LOD) was 0.53 fM. The proposed system was applied to detect miRNA-21 from the urine of acute kidney injury (AKI) patients with good results.

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Data availability

All data in the paper are reliable. Since the majority of the experimental data were obtained from clinical samples of AKI patients, data are not publicly available according to ethical requirements.

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Funding

This project was financially supported by the Basic and Applied Basic Research Foundation of Guangdong Province (No. 2021A1515220136) and the National Natural Science Foundation of China (No. 22174018).

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Author notes

  1. Yunpeng Xu and Jiahui Chen contributed equally to this work.

Authors and Affiliations

  1. Department of Nephrology, People’s Hospital of Bao’an District, Shenzhen, Guangdong, 518000, People’s Republic of China

    Yunpeng Xu, Jiahui Chen, Xiaolu Sui, Yanzi Zhang, Aisha Zhang & Jihong Chen

  2. Bao’an Shenzhen Clinical Medical School of Guangdong Medical University, Shenzhen, Guangdong, 518000, People’s Republic of China

    Yunpeng Xu, Jiahui Chen & Jihong Chen

  3. Ministry of Education Key Laboratory for Analysis Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fujian, Fuzhou, 350116, People’s Republic of China

    Zhenyu Lin

  4. Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics, Institute of Aging Research, Guangdong Medical University, Dongguan, Guangdong, 523808, People’s Republic of China

    Xinguang Liu

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  1. Yunpeng Xu
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Contributions

Yunpeng Xu: conceptualization, investigation, writing – original draft, editing. Jiahui Chen: investigation, writing – original draft. Xiaolu Sui: investigation, data curation. Yanzi Zhang: investigation. Aisha Zhang: investigation. Xinguang Liu: writing – review and editing, supervision. Zhenyu Lin: writing – review and editing, methodology, supervision. Jihong Chen: writing – review and editing, supervision.

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Correspondence to Zhenyu Lin, Xinguang Liu or Jihong Chen.

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Xu, Y., Chen, J., Sui, X. et al. Ultra-sensitive electrochemiluminescent biosensor for miRNA based on CRISPR/Cas13a trans-cleavage-triggered hybridization chain reaction and magnetic-assisted enrichment. Microchim Acta 190, 393 (2023). https://doi.org/10.1007/s00604-023-05962-1

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