Authors
Xinyao Li, Xinyu Gao, Tao Gu, Qiong Li, Li Wang, Fei Deng, Mingyi Guo, Danqun Huo, Changjun Hou
Published in
Analytical chemistry. Jul 09, 2026. Epub Jul 09, 2026.
Abstract
Accurate detection of human epidermal growth factor receptor 2 (HER2) is critical for early breast cancer screening and personalized therapy. This study constructed a target-triggered, hairpin-enhanced CRISPR/Cas12a biosensor named "Tribos" for ultrasensitive HER2 detection. The system integrates an aptamer hairpin switch (HAS), HAS-allosterically triggered rolling circle amplification (RCA), and a hairpin-enhanced CRISPR/Cas12a fluorescence reporter module. Taking advantage of Cas12a's high affinity for stem-loop structures, we designed a double-stem-loop reporter probe (DS-FQ) and validated its trans-cleavage enhancement mechanism via molecular docking. Under optimal conditions, Tribos exhibited a linear range from 10 fg/mL to 10 ng/mL, with a limit of detection as low as 1.08 fg/mL. In clinical validation with 29 breast cancer patients and 13 healthy controls, the sensor achieved a sensitivity of 82.76% and a specificity of 100%, which were highly consistent with clinical diagnoses and ELISA results, and it effectively distinguished different HER2 expression levels. The modular design of Tribos offers a new strategy for high-performance CRISPR diagnostics and lays a foundation for next-generation molecular diagnostic technologies based on nucleic acid conformational regulation.
PMID:
42424600
Bibliographic data and abstract were imported from PubMed on 10 Jul 2026.
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