Lateral flow biosensor-assisted loop-mediated isothermal amplification assay for ultrafast, sensitive detection of African swine fever virus

African swine fever (ASF) is an acute and highly lethal infectious disease caused by African swine fever virus (ASFV). ASF has caused huge economic losses in many countries, and there is still a lack of effective vaccines and treatments. Therefore, the development of rapid and accurate detection methods is crucial for the effective prevention and control of swine fever. Here, we developed a novel loop-mediated isothermal amplification (LAMP) combined with nanoparticle-based lateral flow biosensor (LFB) for rapid and specific ASFV detection (ASFV-LAMP-LFB). The ASFV-LAMP-LFB assay was optimised to allow LAMP-LFB results to be visually verified by LFB within 2-5 min, with the entire detection process completed in 35 min and achieving a sensitivity of 0.04 fg/µL. Concurrently, the specificity of the ASFV-LAMP-LFB assay was evaluated using a range of pathogens, yielding 100% specificity and no cross-reactivity with other porcine pathogens. In addition, the ASFV-LAMP-LFB assay exhibited a 100% degree of sensitivity and specificity in simulated samples. In conclusion, the ASFV-LAMP-LFB assay established in this study is highly sensitive and specific, and can rapidly and accurately identify ASFV. This study offers a technical complement and support for the detection of ASFV in slaughterhouses, farms, and other grassroots units.