Abstract
The present chapter describes a simple and economic indirect enzyme immunoassay (ELISA ) for African swine fever virus (ASFV) antibody detection based on the use of the soluble cytoplasmic fraction of ASFV-infected monkey stable cells (MS). The soluble antigen proteins of ASFV-infected cells are separated by sucrose precipitation centrifugation, and the supernatant above the sucrose layer is used as an ELISA antigen. The test serum sample reacts with the cytoplasmic soluble fraction, and antibodies are detected using a protein A-peroxidase conjugate. This crude antigen is currently recommended as a test reagent in screening and diagnostic tests by the World Organization for Animal Health (OIE).
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References
Gallardo C, Fernández-Pinero J, Arias M (2019) African swine fever (ASF) diagnosis, an essential tool in the epidemiological investigation. Virus Res 271:197676. https://doi.org/10.1016/j.virusres.2019.197676
World Organisation for Animal Health (OIE) (2019) African swine fever. In: Manual of diagnostic tests and vaccines for terrestrial animals 2019, vol 2, Chapter 3.8.2 [cited 2021 March 21]. https://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/3.08.01_ASF.pdf
Gallardo C, Soler A, Nieto R et al (2015) Experimental transmission of African swine fever (ASF) low virulent isolate NH/P68 by surviving pigs. Transbound Emerg Dis 62(6):612–622. https://doi.org/10.1111/tbed.12431
Arias M, Sanchez-Vizcaino JM (2002) African swine fever eradication: the Spanish model. In: Trends in emerging viral infections of swine, 1st edn. Iowa State University Press, pp 133–139. ISBN: 0-8138-0383-7. 2002. (chapter)
Danzetta ML, Marenzoni ML, Iannetti S et al (2020) African swine fever: lessons to learn from past eradication experiences. A systematic review. Front Vet Sci 7:296. https://doi.org/10.3389/fvets.2020.00296
Gallardo C, Nieto R, Soler A et al (2015) Assessment of African swine fever diagnostic techniques as a response to the epidemic outbreaks in eastern European Union countries: how to improve surveillance and control programs. J Clin Microbiol 53(8):2555–2565. https://doi.org/10.1128/JCM.00857-15
Sanchez-Vizcaino JM, Martín L, Ordás A (1979) Adaptación y evaluación del enzimoinmunoensayo para la detección de anticuerpos de peste porcina africana. Laboratorio 67:311–319
Escribano JM, Pastor MJ, Sánchez-Vizcaíno JM (1989) Antibodies to bovine serum albumin in swine sera: implications for false-positive reactions in the serodiagnosis of African swine fever. Am J Vet Res 50(7):1118–1122
Escribano JM, Tabarés E (1987) Proteins specified by African swine fever virus: V. identification of immediate early, early and late proteins. Arch Virol 92(3–4):221–232. https://doi.org/10.1007/BF01317479
Tabarés E, Martinez J, Ruiz Gonzalvo F et al (1980) Proteins specified by African swine fever virus. II Analysis of proteins in infected cells and antigenic properties. Arch Virol 66(2):119–132. https://doi.org/10.1007/BF01314980
Alcaraz C, De Diego M, Pastor MJ et al (1990) Comparison of a radioimmunoprecipitation assay to immunoblotting and ELISA for detection of antibody to African swine fever virus. J Veterinary Diagn Invest 2(3):191–196. https://doi.org/10.1177/104063879000200307
Pastor MJ, Arias M, Escribano JM (1990) Comparison of two antigens for use in an enzyme-linked immunosorbent assay to detect African swine fever antibody. Am J Vet Res 51(10):1540–1543
Gallardo C, Soler A, Nieto R et al (2013) Comparative evaluation of novel African swine fever virus (ASF) antibody detection techniques derived from specific ASF viral genotypes with the OIE internationally prescribed serological tests. Vet Microbiol 162(1):32–43. https://doi.org/10.1016/j.vetmic.2012.08.01
Beltrán-Alcrudo D, Arias M, Gallardo C et al (2017) African swine fever: detection and diagnosis – a manual for veterinarians. FAO animal production and health manual no. 19. Food and agriculture Organization of the United Nations (FAO), Rome. 88 page. Available at http://www.fao.org/3/a-i7228e.pdf
World Organisation for Animal Health (OIE) (2021) Biosafety and biosecurity: Standard for managing biological risk in the veterinary laboratory and animal facilities (version adopted in May 2015). In: Manual of diagnostic tests and vaccines for terrestrial animals 2021 Part. 1 Chapter 1.1.4 [cited 2022 February 22]. https://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/1.01.04_BIOSAFETY_BIOSECURITY.pdf
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Carmina, G., Nieto, R., Arias, M. (2022). African Swine Fever Virus (ASFV) Indirect ELISA Test Based on the Use of the Soluble Cytoplasmic Semi- purified Antigen (ASFV CP-Ag). In: Netherton, C.L. (eds) African Swine Fever Virus. Methods in Molecular Biology, vol 2503. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2333-6_9
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DOI: https://doi.org/10.1007/978-1-0716-2333-6_9
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