Evaluation of the Flashtest multiplex qPCR system for rapid pre-laboratory screening for African swine fever infection

Abstract

Introduction: African swine fever (ASF) is a devastating viral disease of domestic pigs and wild boar. As no safe vaccine is currently available, prevention relies primarily on strict biosecurity measures. Early diagnosis is essential to reduce the risk of ASF spread. Among the available methods, qPCR is considered the gold standard and is recommended by the World Organisation for Animal Health. The Flashtest system is designed to perform the complete workflow, from genetic material extraction to qPCR analysis, enabling rapid (under 1 hour) and accurate detection of various animal diseases in veterinary practice. Material and Methods: In this study, the Flashtest system was evaluated in ASF detection using blood samples collected during ASF outbreaks in 2025 in Poland and samples from experimentally infected pigs. Its performance was compared with a validated commercial qPCR assay employed by the National Reference Laboratory for ASF in Poland. Results: The system achieved up to 97% sensitivity, 100% specificity and reproducibility with a coefficient of variation below 3.5%. The limit of detection was estimated at 10³ TCID₅₀/mL, and a slightly reduced amplification efficiency was observed. Conclusion: These results indicate that the Flashtest system reliably detects ASF in the majority of cases. Nevertheless, confirmatory testing is recommended for samples with low viral loads.