Target-triggered exponential amplification-enhanced CRISPR/Cas12a system with a fluorescence-quenching lateral flow assay for ultrasensitive detection of Salmonella typhimurium.

Authors

Yuanqian Xiong, Li Li, Jun Huang, Xuan-Ang Shen, Jiayi Sun, Yuqian Guo, Daofeng Liu, Tongtong Ma, Xiaolin Huang, Yonghua Xiong

Published in

Biosensors & bioelectronics. Volume 316. Pages 119351. Oct 21, 2026. Epub Oct 21, 2026.

Abstract

The use of the CRISPR/Cas12a system combined with lateral flow assays shows significant promise for point-of-care testing in resource-limited regions. However, the use of isothermal amplification technologies to preenrich target DNA fragments inherently increases the risk of aerosol contamination during strip testing. Here, we introduce a target-triggered exponential amplification-enhanced CRISPR/Cas12a system combined with a fluorescence-quenching lateral flow assay (TEA-FQLFA) for ultrasensitive detection of Salmonella typhimurium. The TEA system converts a single Cas12a recognition event into sequential dual Cas12a activation, initiating a cleavage cascade amplification that achieves a 10⁶-fold signal enhancement without the need for a target DNA preamplification operation. Moreover, hyperbranched gold-coated polymer microspheres (HGPMs), characterized by ultrahigh extinction coefficients and a broad absorption band in the 300-800 nm range, are utilized as fluorescence quenching reporters of the FQLFA for simultaneous colorimetric visualization and fluorescence-based quantitative detection of target DNA concentration. By leveraging the synergistic integration of TEA-mediated signal amplification and HGPM-enabled optical signal enhancement, the TEA-FQLFA system achieves attomolar-level detection of target DNA (0.081 aM) and enables sensitive detection of Salmonella typhimurium in complex food matrices, with detection limits of 7.95 CFU/mL in colorimetric mode and 6.94 CFU/mL in fluorescence mode. In summary, this work establishes a versatile platform that integrates cascade signal amplification with dual-mode lateral-flow for ultrasensitive food safety detection.

PMID:
42913300
Bibliographic data and abstract were imported from PubMed on 24 Oct 2026.

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