High level expression of the major antigenic African swine fever virus proteins p54 and p30 in baculovirus and their potential use as diagnostic reagents

At present, the eradication of African swine fever (ASF) in affected countries is based only on an efficient diagnosis program because of the absence of an available vaccine. The highly antigenic ASF virus proteins p54 and p30, encoded by genes E183L and CP204L respectively, were expressed in baculovirus for diagnostic purposes. A sequence comparison analysis of these genes from different field virus strains which are geographically diverse and isolated in different years, revealed that both genes are completely conserved among the isolates. Partially purified baculovirus-expressed proteins were used in ELISA and Western blot for ASF antibody detection in sera from experimentally inoculated pigs and field sera from ASF innaparent carriers. These comparative analyses showed that p54 presents better reactivity than p30 in Western blot. However, recombinant p30 was more efficient for antibody detection by ELISA, improving the discrimination between positive and negative sera by this technique. These data suggest the convenience of using p30 as ELISA antigen, while p54 should be the selected antigen for ASF virus antibody detection by Western blot. The combined use of both antigens for serodiagnosis of ASF disease will improve the sensitivity of innaparent carriers detection, facilitating also the interpretation of the tests, and eliminating the use of ASF virus in antigen production.

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Main Authors: Oviedo, J. M., Rodriguez, F., Gómez-Puertas, P., Brun Torres, Alejandro, Gómez, N., Alonso, C., Escribano, J. M.
Format: artículo biblioteca
Language:English
Published: Elsevier 1997
Subjects:African swine fever, ELISA, Serodiagnosis, Antigen production,
Online Access:http://hdl.handle.net/20.500.12792/3260
http://hdl.handle.net/10261/292817
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spelling dig-inia-es-10261-2928172023-02-20T07:32:45Z High level expression of the major antigenic African swine fever virus proteins p54 and p30 in baculovirus and their potential use as diagnostic reagents Oviedo, J. M. Rodriguez, F. Gómez-Puertas, P. Brun Torres, Alejandro Gómez, N. Alonso, C. Escribano, J. M. African swine fever ELISA Serodiagnosis Antigen production At present, the eradication of African swine fever (ASF) in affected countries is based only on an efficient diagnosis program because of the absence of an available vaccine. The highly antigenic ASF virus proteins p54 and p30, encoded by genes E183L and CP204L respectively, were expressed in baculovirus for diagnostic purposes. A sequence comparison analysis of these genes from different field virus strains which are geographically diverse and isolated in different years, revealed that both genes are completely conserved among the isolates. Partially purified baculovirus-expressed proteins were used in ELISA and Western blot for ASF antibody detection in sera from experimentally inoculated pigs and field sera from ASF innaparent carriers. These comparative analyses showed that p54 presents better reactivity than p30 in Western blot. However, recombinant p30 was more efficient for antibody detection by ELISA, improving the discrimination between positive and negative sera by this technique. These data suggest the convenience of using p30 as ELISA antigen, while p54 should be the selected antigen for ASF virus antibody detection by Western blot. The combined use of both antigens for serodiagnosis of ASF disease will improve the sensitivity of innaparent carriers detection, facilitating also the interpretation of the tests, and eliminating the use of ASF virus in antigen production. 2023-02-20T07:32:45Z 2023-02-20T07:32:45Z 1997 artículo Journal of Virological Methods 64(1): 27-35 (1997) 0166-0934 http://hdl.handle.net/20.500.12792/3260 http://hdl.handle.net/10261/292817 10.1016/S0166-0934(96)02140-4 en none Elsevier
institution INIA ES
collection DSpace
country España
countrycode ES
component Bibliográfico
access En linea
databasecode dig-inia-es
tag biblioteca
region Europa del Sur
libraryname Biblioteca del INIA España
language English
topic African swine fever
ELISA
Serodiagnosis
Antigen production
African swine fever
ELISA
Serodiagnosis
Antigen production
spellingShingle African swine fever
ELISA
Serodiagnosis
Antigen production
African swine fever
ELISA
Serodiagnosis
Antigen production
Oviedo, J. M.
Rodriguez, F.
Gómez-Puertas, P.
Brun Torres, Alejandro
Gómez, N.
Alonso, C.
Escribano, J. M.
High level expression of the major antigenic African swine fever virus proteins p54 and p30 in baculovirus and their potential use as diagnostic reagents
description At present, the eradication of African swine fever (ASF) in affected countries is based only on an efficient diagnosis program because of the absence of an available vaccine. The highly antigenic ASF virus proteins p54 and p30, encoded by genes E183L and CP204L respectively, were expressed in baculovirus for diagnostic purposes. A sequence comparison analysis of these genes from different field virus strains which are geographically diverse and isolated in different years, revealed that both genes are completely conserved among the isolates. Partially purified baculovirus-expressed proteins were used in ELISA and Western blot for ASF antibody detection in sera from experimentally inoculated pigs and field sera from ASF innaparent carriers. These comparative analyses showed that p54 presents better reactivity than p30 in Western blot. However, recombinant p30 was more efficient for antibody detection by ELISA, improving the discrimination between positive and negative sera by this technique. These data suggest the convenience of using p30 as ELISA antigen, while p54 should be the selected antigen for ASF virus antibody detection by Western blot. The combined use of both antigens for serodiagnosis of ASF disease will improve the sensitivity of innaparent carriers detection, facilitating also the interpretation of the tests, and eliminating the use of ASF virus in antigen production.
format artículo
topic_facet African swine fever
ELISA
Serodiagnosis
Antigen production
author Oviedo, J. M.
Rodriguez, F.
Gómez-Puertas, P.
Brun Torres, Alejandro
Gómez, N.
Alonso, C.
Escribano, J. M.
author_facet Oviedo, J. M.
Rodriguez, F.
Gómez-Puertas, P.
Brun Torres, Alejandro
Gómez, N.
Alonso, C.
Escribano, J. M.
author_sort Oviedo, J. M.
title High level expression of the major antigenic African swine fever virus proteins p54 and p30 in baculovirus and their potential use as diagnostic reagents
title_short High level expression of the major antigenic African swine fever virus proteins p54 and p30 in baculovirus and their potential use as diagnostic reagents
title_full High level expression of the major antigenic African swine fever virus proteins p54 and p30 in baculovirus and their potential use as diagnostic reagents
title_fullStr High level expression of the major antigenic African swine fever virus proteins p54 and p30 in baculovirus and their potential use as diagnostic reagents
title_full_unstemmed High level expression of the major antigenic African swine fever virus proteins p54 and p30 in baculovirus and their potential use as diagnostic reagents
title_sort high level expression of the major antigenic african swine fever virus proteins p54 and p30 in baculovirus and their potential use as diagnostic reagents
publisher Elsevier
publishDate 1997
url http://hdl.handle.net/20.500.12792/3260
http://hdl.handle.net/10261/292817
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