Conference in 118th Annual Meeting United States Animal Health Association and 57th Annual Conference American Association of Veterinary Laboratory Diagnosticians

October 16^th, 2014

Gimenez-Lirola LG., Zimmerman JJ., Mur L., Rivera B., Sanchez-Vizcaino JM., Lizano S., Goodell CK., Rowland R., Mogler MA., Harris DL., Gallardo C. and Arias M.

African swine fever (ASF) is a devastating, highly contagious disease classified as a Foreign Animal Disease (FAD) in the U.S. Serology has been widely used in ASFV control programs in the Iberian Peninsula and Sardinia as a tool for the detection of ASFV carrier animals. Among ASFV proteins considered to be candidate antigens for serological tests, structural proteins p30, p54, and p72 are the best described, most highly studied, and most widely used in commercial ASFV serum antibody ELISAs1.
Serum and oral fluid antibody-positive samples were generated by experimental inoculation of 17 pigs with an attenuated ASFV isolate (NHV) that produces chronic infection. Oral fluid and serum samples were sequentially collected over days post inoculation (DPI 0, 6, 12, 15, 19, 26, 33, 40, 47, 54, and 61) using methods previously described2. The performance of the
optimized ELISA was also evaluated using serum (n = 200) and oral fluid (n = 200) samples from animals (n = 400) known to be free of ASFV infection.
The antigen used in the ELISA was selected by evaluating the serum antibody response of ASFV-infected pigs against three recombinant antigens (rp30, rp54, rp72) using a multiplex fluorescent microbead-based immunoassay (FMIA; Luminex® Corporation). Antibody was detected at 6 DPI against p72 (11%) and at 12 DPI for both p30 (100%), and p54 (89%). All pigs (100%) were positive at DPI 12 for p30, at DPI 15 for p54, and at DPI 19 for p72.
Recombinant p30 was selected as antigen target for subsequent development of an antibody ELISA.
ASFV rp30 antibody ELISA was able to detect ASFV antibodies by DPI 12 in both serum and oral fluid specimens run on the same plate simultaneously. The evaluation of known ASFV negative field samples showed specificities of 99.5% and 100% for serum and oral fluid samples, respectively. Given the increased surveillance efficiency provided by oral fluid sampling and the ability to corroborate results using serum samples, the ASFV rp30 antibody
would be a highly useful under conditions that warrant ASFV surveillance

	Veterinary Medical Research Institute (VMRI). Hungarian Academy of Sciences.
	Veterinary Diagnostic and Production Animal Medicine (VDPAM). Iowa State University (ISU).
	Servicio de Inmunología Viral y Medicina Preventiva (SUAT). Centro de Vigilancia Sanitaria Veterinaria (VISAVET). Universidad Complutense (UCM).
	Departamento de Sanidad Animal. Facultad de Veterinaria. Universidad Complutense (UCM).
	Idexx Laboratories (IDEXX).
	Department of Diagnostic Medicine/Pathobiology. College of Veterinary Medicine. Kansas State University (KSU).
	Harrisvaccines, Inc.
	Centro de Investigación en Sanidad Animal (CISA). Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA). Ministerio de Ciencia e Innovación.