tetano
Editor, Senior Moderator
Virol J
. 2026 Jan 25.
doi: 10.1186/s12985-026-03085-6. Online ahead of print.
Establishment of a CRISPR/Cas12a/13a-driven dual-detection platform for rapid diagnosis of swine influenza virus and porcine reproductive and respiratory syndrome virus infection
Shuchang Guo[SUP] 1 2 [/SUP], Shiyuchen Zhao[SUP] 1 2 [/SUP], Siqi Tang[SUP] 1 2 [/SUP], Haoyu Leng[SUP] 1 2 [/SUP], Yanan Wu[SUP] 1 2 [/SUP], Wen Li[SUP] 1 2 [/SUP], Shiqi Xing[SUP] 1 2 [/SUP], Yali Feng[SUP] 1 2 [/SUP], Ying Zhang[SUP] 3 4 [/SUP]
Affiliations
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.
Keywords: CRISPR-Cas12a/13a; Lateral flow dipstick (LFD); Porcine reproductive and respiratory syndrome virus (PRRSV); Reverse transcription loop-mediated isothermal amplification (RT-LAMP); Swine influenza virus (SIV).
. 2026 Jan 25.
doi: 10.1186/s12985-026-03085-6. Online ahead of print.
Establishment of a CRISPR/Cas12a/13a-driven dual-detection platform for rapid diagnosis of swine influenza virus and porcine reproductive and respiratory syndrome virus infection
Shuchang Guo[SUP] 1 2 [/SUP], Shiyuchen Zhao[SUP] 1 2 [/SUP], Siqi Tang[SUP] 1 2 [/SUP], Haoyu Leng[SUP] 1 2 [/SUP], Yanan Wu[SUP] 1 2 [/SUP], Wen Li[SUP] 1 2 [/SUP], Shiqi Xing[SUP] 1 2 [/SUP], Yali Feng[SUP] 1 2 [/SUP], Ying Zhang[SUP] 3 4 [/SUP]
Affiliations
- PMID: 41582136
- DOI: 10.1186/s12985-026-03085-6
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.
Keywords: CRISPR-Cas12a/13a; Lateral flow dipstick (LFD); Porcine reproductive and respiratory syndrome virus (PRRSV); Reverse transcription loop-mediated isothermal amplification (RT-LAMP); Swine influenza virus (SIV).