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A CRISPR/Cas12a-based aptasensor enhanced by functionalized AuNPs for sensitive full-range C-reactive protein

Siyue Wang1, Xiaotian Guan1, Shuqing Sun1

  • 1Shenzhen International Graduate School, Tsinghua University, Shenzhen, 518055, China. sun.shuqing@sz.tsinghua.edu.cn.

Journal of Materials Chemistry. B
|October 31, 2025
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Summary

A new fluorescence aptasensor, AuCA, offers sensitive and broad-range detection of C-reactive protein (CRP). This novel method, utilizing CRISPR-Cas12a technology, shows promise for accurate clinical diagnostics of inflammatory and cardiovascular conditions.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • C-reactive protein (CRP) is a key biomarker for inflammation, infection, and cardiovascular risk.
  • Existing CRP detection methods face challenges in balancing sensitivity and dynamic range.
  • Accurate and robust CRP quantification is crucial for clinical diagnostics.

Purpose of the Study:

  • To develop and validate a novel, highly sensitive, and broad-dynamic-range aptasensor for CRP detection.
  • To integrate aptamers, magnetic beads, gold nanoparticles, and CRISPR-Cas12a for enhanced CRP quantification.
  • To assess the performance of the developed aptasensor in clinical human plasma samples.

Main Methods:

  • Development of the AuNP-enhanced CRISPR Aptasensor (AuCA) integrating aptamers, magnetic beads, and gold nanoparticles.
  • Utilizing the CRISPR/Cas12a system for signal amplification via trans-cleavage of fluorescent reporters.
  • Validation of the AuCA platform using standard and clinical human plasma samples, comparing results with a commercial assay.

Main Results:

  • The AuCA platform achieved a low detection limit of 60 ng mL⁻¹ and a broad quantifiable range of 0.1–150 μg mL⁻¹.
  • Demonstrated excellent specificity and resistance to biological interference in complex matrices.
  • Showed strong concordance with a commercial immunoturbidimetric assay for CRP detection in human plasma.
  • Enabled simultaneous detection of standard and hypersensitive CRP (hsCRP) for full-range CRP (frCRP) analysis.

Conclusions:

  • The AuCA aptasensor provides a robust and sensitive method for full-range CRP quantification.
  • This technology holds significant potential for improved clinical diagnostics of inflammatory and cardiovascular diseases.
  • The developed platform offers a promising alternative to conventional CRP detection techniques.