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- Title
SERS-based CRISPR/Cas12a assays for protein biomarker prostate-specific antigen detection: SERS-based CRISPR/Cas12a assays for protein biomarker prostate-specific antigen detection: P. Teng et al.
- Authors
Teng, Peijun; Gao, Zhixing; Quan, Qiang; He, Guangbo; Song, Qifang; Zhang, Xiaoli; Xiao, Wei; Zhao, Jianfu; Cao, Donglin; Liang, Jiajie; Tang, Yong
- Abstract
Sensitive and accurate detection of protein biomarkers is crucial for disease diagnosis, especially for early diagnosis. Here, we describe surface-enhanced Raman scattering (SERS)-based CRISPR/Cas12a assays (S-CRISPR) for protein biomarker detection. Firstly, an S-CRISPR-driven enzyme-linked immunosorbent assay (S-CasLISA) was developed utilizing a capture antibody coated on a microplate to recognize the target and a detection antibody labeled with active DNA to trigger the activity of CRISPR/Cas12a. With this assay, we achieved detection of prostate-specific antigen (PSA) as models at the picogram level. The limit of detection (LoD) of S-CasLISA was 0.17 pg mL−1 and in the range of 0.1 pg mL−1 to 10 ng mL−1. Further, we applied aptamer to S-CRISPR (S-Apt-CRISPR), combining the high sensitivity of SERS with the high selectivity of aptamers, while simplifying the operation process of CRISPR detection of protein biomarkers. The proposed S-Apt-CRISPR also could detect picogram-level PSA and without repeated washing steps. The LoD of S-Apt-CRISPR was 0.35 pg mL−1 and in the range of 0.1 pg mL−1 to 10 ng mL−1. Both SERS-based CRISPR/Cas12a assays were validated with clinical samples and demonstrated accuracy consistent with the chemiluminescence immunoassay. The introduction of the CRISPR/Cas12a system with SERS has the effect of improving the analytical capabilities of the system, thereby broadening and facilitating its application in the analysis of sensitive and accurate protein biomarkers.
- Subjects
CRISPRS; ENZYME-linked immunosorbent assay; PROSTATE-specific antigen; CHEMILUMINESCENCE immunoassay; BIOMARKERS
- Publication
Analytical & Bioanalytical Chemistry, 2025, Vol 417, Issue 3, p573
- ISSN
1618-2642
- Publication type
Academic Journal
- DOI
10.1007/s00216-024-05663-5