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

Oct 31, 2025Journal of materials chemistry. B

Sensitive detection of C-reactive protein using a CRISPR-based sensor improved with special gold nanoparticles

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Abstract

The AuCA platform achieved a low detection limit of 60 ng/mL for C-reactive protein (CRP).

Aptamer-functionalized magnetic beads selectively captured CRP, enhancing detection capabilities.
Gold nanoparticles modified with an activation sequence facilitated efficient signal transduction.
Upon CRP binding, the system activated a Cas12a complex, leading to amplified fluorescent signals.
AuCA demonstrated a quantifiable range of 0.1-150 μg/mL and maintained excellent specificity.
Clinical application in human plasma yielded results closely aligned with a commercial assay.

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C-reactive protein (CRP) is a well-established biomarker whose plasma levels increase significantly during inflammatory and infectious episodes. It plays a vital role in the diagnosis of bacterial infections and autoimmune diseases, and cardiovascular risk assessment. However, conventional detection techniques often struggle to balance high sensitivity with a broad dynamic range. Here, we developed and validated a novel fluorescence-based aptasensor named AuCA (AuNP-enhanced CRISPR Aptasensor), which integrates nucleic acid aptamers, magnetic beads (MBs), gold nanoparticles (AuNPs), and the CRISPR/Cas12a system for robust CRP quantification. In this system, MBs functionalized with aptamer 1 selectively captured the target protein, while AuNPs co-modified with aptamer 2 and a Cas12a activation sequence (Trigger) enabled efficient signal transduction. Upon target binding, the Trigger activated the Cas12a/crRNA complex, initiating trans-cleavage of fluorescent reporters and producing a markedly amplified signal. The optimized AuCA platform achieved a low detection limit of 60 ng mLand a quantifiable range of 0.1-150 μg mL. It exhibited excellent specificity and resistance to biological interference, ensuring reliable measurements even in complex sample matrices. When applied to clinical human plasma, AuCA demonstrated results that are in strong concordance with results from a commercial immunoturbidimetric assay. AuCA allowed the simultaneous detection of both standard and hypersensitive CRP (hsCRP), supporting comprehensive full-range CRP (frCRP) analysis with strong potential for clinical applications. -1 -1

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

Abstract

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