High-Accuracy Serodiagnosis of African Swine Fever Using P72 and P30-Based Lateral Flow Assays: A Validation Study...

African Swine Fever (ASF) control is severely hampered by the reliance on slow, laboratory-bound diagnostics. While rapid, field-deployable lateral flow assays (LFAs) are urgently needed, the comparative performance of key single-antigen targets remains poorly characterized. This study aimed to develop and systematically evaluate the diagnostic performance of three in-house single-antigen LFAs targeting ASF virus P30, P54, and P72, using swine field samples from Thailand, including a panel of 143 quantitative polymerase chain reaction-negative swine serum samples. The performance of each LFA was compared against a commercial multi-antigen (P32/P62/P72) indirect ELISA, which served as the reference standard, classifying 64 samples as positive and 79 as negative. The P72-based LFA demonstrated perfect diagnostic performance (100% sensitivity, 100% specificity) and perfect agreement (κ = 1.0) with the enzyme-linked immunosorbent assay (ELISA). Similarly, the P30 LFA demonstrated high performance (100% sensitivity, 98.7% specificity) with 'Almost Perfect' agreement (κ = 0.9859). In contrast, the P54 LFA was unsuitable, achieving 100% sensitivity but unacceptably low specificity (88.6%) due to a high rate of false positives. Overall, the single-antigen P72 and P30 LFAs demonstrated excellent concordance with the multi-antigen ELISA, supporting their reliable for detecting antibodies against ASFV. Although these assays do not replace molecular methods for acute infection detection, they represent valuable complementary tools for serosurveillance.