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

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.