African swine fever (ASF) is a contagious disease of wild boar and domestic pigs notifiable to the World Organisation for Animal Health due to its high socio-economic impact. ASF is caused by the complex ASF virus (ASFV), and it can present different clinical manifestations that can be confused with other diseases; for this reason, laboratory testing is necessary for the proper diagnosis of clinically suspected animals. Despite the efforts put into it over decades, no treatment or safe vaccine is globally available, and disease control is based on early diagnosis and the implementation of strict biosecurity measures. In this context, rapid tests have the potential to accelerate and facilitate the identification of infected animals by giving fast on-site results. In this work, we improved the available point-of-care assays for the diagnosis of the disease by the development of a more specific antigen test and a more sensitive antibody test. This antibody detection test allowed for the earlier detection of infected animals than two commercial indirect ELISAs (statistically significant). Moreover, we developed a combined dual rapid test, unifying, in the same cassette, an antigen detection strip and an antibody detection strip. In this study, we confirmed that this combo approach is a useful tool for implementing rapid tests in the field since it increases the percentage of positive samples detected, even when PCR turns negative, while maintaining a good specificity.

Department of Virology State Veterinary Institute Jihlava Rantirovska 93 20 Horni Kosov 586 01 Jihlava Czech Republic

European Union Reference Laboratory for African Swine Fever Centro de Investigación en Sanidad Animal Carretera Algete El Casar de Talamanca Km 8 1 28130 Madrid Spain

Gold Standard Diagnostics Madrid Calle de los Hermanos García Noblejas 39 28037 Madrid Spain

Institute of Food Safety Animal Health and Environment BIOR Lejupes Street 3 LV 1076 Riga Latvia

Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche Togo Rosati Via Salvemini 1 06126 Perugia Italy

National Food and Veterinary Risk Assessment Institute J Kairiūkščio g 10 LT 08409 Vilnius Lithuania

Virology Department Institute of Veterinary Medicine of Serbia Janisa Janulisa 14 11000 Belgrade Serbia

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