REVIEW ARTICLE African Swine Fever Diagnosis Adapted to Tropical Conditions by the Use of Dried-blood Filter Papers T. Randriamparany 1, *, K. V. Kouakou 2, *, V. Michaud 3,4 , J. Fern andez-Pinero 5 , C. Gallardo 5 , M.-F. Le Potier 6 , R. Rabenarivahiny 1 , E. Couacy-Hymann 7 , M. Raherimandimby 8 and E. Albina 4,9 1 Laboratoire National de Diagnostic V et erinaire, Antananarivo, Madagascar 2 Laboratoire National d’Appui au D eveloppement Agricole, Bingerville, C^ ote-d’Ivoire 3 CIRAD, UMR CMAEE, Montpellier, France 4 INRA, UMR1309 CMAEE, Montpellier, France 5 Centro de Investigacion en Sanidad Animal (CISA-INIA), Valdeolmos, Spain 6 Anses, Laboratoire de Ploufragan, Unit e Virologie Immunologie Porcines, Ploufragan, France 7 LANADA/Laboratoire Central de Pathologie Animale, Bingerville, C^ ote-d’Ivoire 8 Universit e d’Antananarivo, Antananarivo, Madagascar 9 CIRAD, UMR CMAEE, Petit-Bourg, Guadeloupe, France Keywords: African swine fever; sample collection; filter paper; diagnosis; ELISA; PCR Correspondence: E. Albina. CIRAD, UMR CMAEE, Domaine Duclos, 97170 Petit Bourg, Guadeloupe, France. Tel.: +33 59025431; Fax: +33 590940396; E-mail: emmanuel.albina@cirad.fr *These two authors contributed equally to this work. Received for publication July 4, 2014 doi:10.1111/tbed.12295 Summary The performance of Whatman 3-MM filter papers for the collection, drying, ship- ment and long-term storage of blood at ambient temperature, and for the detec- tion of African swine fever virus and antibodies was assessed. Conventional and real-time PCR, viral isolation and antibody detection by ELISA were performed on paired samples (blood/tissue versus dried-blood 3-MM filter papers) collected from experimentally infected pigs and from farm pigs in Madagascar and C^ ote d’Ivoire. 3-MM filter papers were used directly in the conventional and real-time PCR without previous extraction of nucleic acids. Tests that performed better with 3-MM filter papers were in descending order: virus isolation, real-time UPL PCR and conventional PCR. The analytical sensitivity of real-time UPL PCR on filter papers was similar to conventional testing (virus isolation or conventional PCR) on organs or blood. In addition, blood-dried filter papers were tested in ELISA for antibody detection and the observed sensitivity was very close to con- ventional detection on serum samples and gave comparable results. Filter papers were stored up to 9 months at 20–25°C and for 2 months at 37°C without signifi- cant loss of sensitivity for virus genome detection. All tests on 3-MM filter papers had 100% specificity compared to the gold standards. Whatman 3-MM filter papers have the advantage of being cheap and of preserving virus viability for future virus isolation and characterization. In this study, Whatman 3-MM filter papers proved to be a suitable support for the collection, storage and use of blood in remote areas of tropical countries without the need for a cold chain and thus provide new possibilities for antibody testing and virus isolation. Introduction African swine fever virus (ASFV) is a large enveloped DNA virus (Dixon et al., 2005) that belongs to the family Asfar- viridae, genus Asfivirus. It is the only known DNA arbovi- rus. African swine fever virus infects members of the vertebrate family Suidae (domestic and feral pigs, wild boars, bush pigs, warthogs and the giant forest hog) and some Argasid ticks (Ornithodoros complex). It is one of the most fatal diseases of domestic pigs and wild boars and the most serious transboundary pig disease that could spread rapidly and have crippling socio-economic © 2014 Blackwell Verlag GmbH • Transboundary and Emerging Diseases. 1 Transboundary and Emerging Diseases