Virology journalJan 25, 2026

Rapid detection of swine flu and pig reproductive respiratory syndrome using a CRISPR-based dual test

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗OA ↗

Updated Jan 26, 2026

Abstract

The dual-detection platform achieved detection limits of 5 copies/µL for swine influenza virus (SIV) and 2 copies/µL for porcine reproductive and respiratory syndrome virus (PRRSV).

The assay integrates reverse transcription loop-mediated isothermal amplification with CRISPR technology for enhanced detection.
A 20-minute amplification process at 40 °C is followed by a 5-minute readout using lateral flow dipstick visualization.
High specificity was observed against other common swine pathogens during testing.
Preliminary validation with respiratory infection samples showed high concordance with existing reference methods.

Simplified

BACKGROUND: Swine influenza virus (SIV) and porcine reproductive and respiratory syndrome virus (PRRSV) are leading pathogens in pigs, whose co-infections exacerbate disease severity. Current diagnostics like RT-PCR lack suitability for rapid, on-site use, while CRISPR-based systems face challenges in convenient multiplex detection.

RESULTS: We developed an RT-LAMP-CRISPR-Cas12a/13a-LFD dual-detection platform that integrates reverse transcription loop-mediated isothermal amplification (RT-LAMP) with the orthogonal trans-cleavage activities of CRISPR-Cas12a and Cas13a, followed by lateral flow dipstick (LFD) visualization. This assay achieved detection limits of 5 copies/µL for SIV and 2 copies/µL for PRRSV, and exhibited high specificity against other common swine pathogens. The entire process, including a 20-minute amplification at 40 °C and 5-minute LFD readout, enables rapid and visual diagnosis. A preliminary validation was conducted using respiratory infection samples, demonstrating high concordance with reference methods and specificity against non-target pathogens.

CONCLUSIONS: The RT-LAMP-CRISPR-Cas12a/13a-LFD assay provides a sensitive, specific, and potentially field-adaptable tool for the simultaneous detection of SIV and PRRSV. It is ideally suited for early screening and precise control of these pathogens in resource-limited settings.

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Competing interests

Declarations. Ethics approval and consent to participate: This work complied with all relevant ethical guidelines and did not involve the use of experimental animals. All samples were exclusively sourced from existing surveillance infrastructure; specifically, porcine swabs were collected as part of routine pathogen monitoring programs in multiple swine farms and supplied by the farms’ own diagnostic laboratories. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests.

PubMed

Rapid detection of swine flu and pig reproductive respiratory syndrome using a CRISPR-based dual test

Abstract

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Funding

Competing interests

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