Serum-compatible aptamer-regulated RCA-CRISPR/Cas12a assay for fluorescence and lateral flow detection of myeloperoxidase.

Authors

Yingying Cao, Mengchen Duan, Lizhen Wei, Yisheng Chen, Fei Liu, Jiawen Bai, Yi Huang, Shenghang Zhang

Published in

Mikrochimica acta. Volume 193. Issue 10. Oct 01, 2026. Epub Oct 01, 2026.

Abstract

Myeloperoxidase (MPO), a neutrophil-derived heme peroxidase involved in oxidative inflammatory responses, has been investigated as a biomarker for cardiovascular, hematological, and other inflammation-associated diseases. In this study, an aptamer-regulated rolling circle amplification (RCA)-CRISPR/Cas12a fluorescence assay was established for MPO detection in serum samples. Two aptamers, Apt-02 and Apt-14, were obtained with dissociation constants (K_D) of 57.76 and 56.88 nM, respectively, using MPO-depleted human serum as the counter-selection matrix during MB-SELEX. Apt-14 exhibited relatively good stability in serum and was therefore used in the assay development. Upon MPO binding, cDNA was released from the aptamer/cDNA duplex, initiating RCA and subsequently activating Cas12a-mediated trans-cleavage to amplify the fluorescence signal. Under the optimized conditions, the assay achieved a detection limit of 0.422 ng/mL with good selectivity. It also showed limited interference from the tested cardiovascular disease-related proteins and provided MPO levels in clinical serum samples that were consistent with ELISA results. The assay was also adapted to a lateral-flow strip using a biotin/FAM-labeled ssDNA reporter, enabling visual detection and reader-assisted semi-quantitative analysis.

PMID:
42821128
Bibliographic data and abstract were imported from PubMed on 02 Oct 2026.

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