Abstract
The immunoperoxidase technique (IPT) is an immunocytochemistry technique on fixed infected cells to determine the antibody-antigen complex formation through the action of the peroxidase enzyme. In this procedure, African green monkey established cell lines, such as Vero or MS, are infected with African swine fever virus (ASFV)-adapted isolates to these cell cultures. The infected cells are fixed and then used as antigens to determine the presence of the specific antibodies against ASFV in samples of swine origin. IPT has many advantages for the serological diagnosis of ASF. Firstly, IPT has a higher sensitivity than ELISA or immunoblot assay (IB) for the detection of antibodies against ASFV. Secondly, it has greater versatility to analyze any type of clinical sample of porcine origin and to conduct research studies on the epidemiological situation, through the presence of antibody titers in exudates from organs and tissues. Thirdly, the preparation of IPT plates with cells infected with ASFV is simple, although laborious, that can be stored for a long time and allow the analysis of a large number of samples.
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References
Fenner F, Bachmann PA, Gibbs EPI, Murphy FA, Atuddert MJ, White DO (1987) Laboratory diagnosis of viral diseases. Veter Virol 1987:237–264. https://doi.org/10.1016/B978-0-12-253055-5.50017-7. Epub 2014 Jun 27. PMCID: PMC7173550
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, Tizzani P, Calistri P, Feliziani F (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
Sanchez-Vizcaino JM (1986) African swine fever diagnosis. In: Becker J (ed) African swine fever. Martinus Nijhoff Publishing, Boston, pp 63–71
Bool PH, Ordas A, Sanchez BC (1969) El diagnostico de la peste porcina africana por immunofluorescencia. Bull Off Int Epizoot 72:819–939
Bool PH, Ordas A, Sanchez BC (1970) Le diagnostic par immunofluorescence de la peste porcine africaine. Riv Patronato Biol Anim 14:115–132
Tessler J, Hess WR, Pan IC et al (1974) Immunofluorescence plaque assay for African swine fever virus. Can J Comp Med 38(4):443–447
Pan IC, Huang TS, Hess WR (1982) New method of antibody detection by indirect immunoperoxidase plaque staining for serodiagnosis of African swine fever. J Clin Microbiol 16(4):650–655. https://doi.org/10.1128/JCM.16.4.650-655.1982
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.2012.08.011
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
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
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
Carrascosa AL, Bustos MJ, de Leon P (2011) Methods for growing and titrating African swine fever virus: field and laboratory samples. Curr Protoc Cell Biol Chapter 26:Unit 26.14. https://doi.org/10.1002/0471143030.cb2614s53
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). Indirect Immunoperoxidase Test (IPT) for Detection of Antibodies Against African Swine Fever Virus (ASFV) on African Green Monkey Cell Lines (Vero, MS). 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_10
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DOI: https://doi.org/10.1007/978-1-0716-2333-6_10
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