SERS-based CRISPR/Cas12a assays for protein biomarker prostate-specific antigen detection

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Molecular Biology

Background:

• Sensitive and accurate protein biomarker detection is vital for early disease diagnosis.
• Current methods may lack sensitivity or require complex procedures.
• CRISPR/Cas12a systems offer precise molecular detection capabilities.

Purpose of the Study:

• To develop novel surface-enhanced Raman scattering (SERS)-based CRISPR/Cas12a assays for sensitive protein biomarker detection.
• To establish SERS-CRISPR assays for picogram-level detection of prostate-specific antigen (PSA).
• To evaluate the clinical applicability and accuracy of the developed SERS-CRISPR assays.

Main Methods:

• Development of SERS-CRISPR-driven enzyme-linked immunosorbent assay (S-CasLISA) using capture antibodies and DNA-labeled detection antibodies.
• Integration of aptamers with SERS-CRISPR (S-Apt-CRISPR) for simplified, highly selective protein detection.
• Utilizing SERS for signal amplification and CRISPR/Cas12a for target recognition and collateral cleavage.

Main Results:

• S-CasLISA achieved a limit of detection (LoD) of 0.17 pg mL⁻¹ for PSA, with a dynamic range of 0.1-10 ng mL⁻¹.
• S-Apt-CRISPR demonstrated an LoD of 0.35 pg mL⁻¹ for PSA, simplifying the assay by eliminating washing steps.
• Both SERS-CRISPR assays showed accuracy comparable to chemiluminescence immunoassays when validated with clinical samples.

Conclusions:

• SERS-based CRISPR/Cas12a assays significantly enhance analytical capabilities for protein biomarker detection.
• These assays offer high sensitivity and accuracy, suitable for early disease diagnosis.
• The developed SERS-CRISPR platforms broaden the application of sensitive and accurate protein analysis.