Rapid and ultra-sensitive detection of African swine fever virus antibody on site using QDM based-ASFV immunosensor...

African swine fever (ASF) is a swine viral disease that could cause highly contagious and extremely high mortality, causing huge economic losses to the pig industry. As there is currently no vaccine and effective treatment methods. Therefore, early monitoring is one of the most important solutions to prevent and control ASF. In this study, the dual QDM recombinant virus protein 30 and 54 (P30 and P54) probes and pre-incubation in vitro were proposed for the first time as QDM based-ASFV immunosensor (QAIS) for the ultra-sensitive quantitative detection of ASFV antibodies in serum. In the range from serum dilution of 1:1000 to 1:64000, it showed a good linear relationship (R² = 0.9947), and its detection sensitivity was 1:64000 dilution. Compared with commercial enzyme-linked immunosorbent assay (ELISA) and colloidal gold immunochromatographic strip (CGICS), its detection sensitivity was improved by at least one order of magnitude and four orders of magnitude respectively. In addition, the whole ASFV antibody screening test can be completed in 25 min with simple operation. The performance and practicability of the established QAIS sensor have been verified by ASF-ELISA kit, and its coincidence rate was as high as 98.7% in 151 clinical samples. We firmly believe that the proposed QAIS sensor could potentially be applied to point-of-care testing (POCT) for quantitative ASFV antibody in pig farms.