A triplex-readout CRISPR-Cas12a multimodal biosensing platform for point-of-care detection of avian influenza H5N1.

Publication date: Aug 01, 2026

The highly pathogenic avian influenza A (H5N1) virus poses a significant zoonotic threat to public and animal health. Conventional detection methods often face limitations in complexity, time, and equipment requirements. In this study, we report a rapid and lightweight-equipment triplex diagnostic platform for H5N1 detection, integrating locked nucleic acid (LNA)-assisted target recognition, toehold-mediated strand displacement for signal amplification, and the high specificity of the CRISPR-Cas12a system. The developed assay achieved a detection limit of 3. 7 cD7 10^2 copies/μL for H5N1 pseudovirus RNA within a 60-minute core detection process, and exhibited excellent specificity without cross-reactivity to other influenza subtypes or coronaviruses. Moreover, it successfully identified H5N1 in clinical swab samples, yielding consistent results across three independent readout formats: fluorescence, lateral flow strip, and a portable glucose meter. With advantages of low cost, rapid operation, and multimodal verification capability, this diagnostic system is well suited for point-of-care screening in resource-limited settings and represents a promising tool for frontline outbreak response.

Concepts Keywords
Avian Animals
Coronaviruses Avian influenza
Crispr Biosensing Techniques
Triplex CRISPR-Cas Systems
Zoonotic CRISPR-Cas12a
Glucometer
Humans
Influenza in Birds
Locked nucleic acid
Multimodal detection
Point-of-Care Systems
Rapid Diagnostic Tests
RNA, Viral
RNA, Viral

Semantics

Type Source Name
disease MESH avian influenza
disease MESH face
pathway REACTOME Signal amplification
disease MESH influenza
drug DRUGBANK Dextrose unspecified form
disease MESH Cas
drug DRUGBANK Influenza A virus

Original Article

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