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

被引:68
作者
Oviedo, JM [1 ]
Rodriguez, F [1 ]
GomezPuertas, P [1 ]
Brun, A [1 ]
Gomez, N [1 ]
Alonso, C [1 ]
Escribano, JM [1 ]
机构
[1] CTR INVEST SANIDAD ANIM,INIA,MADRID 28130,SPAIN
关键词
African swine fever; ELISA; serodiagnosis; antigen production;
D O I
10.1016/S0166-0934(96)02140-4
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. Copyright (C) 1997 Elsevier Science B.V.
引用
收藏
页码:27 / 35
页数:9
相关论文
共 21 条
[1]   CHARACTERIZATION OF P-30, A HIGHLY ANTIGENIC MEMBRANE AND SECRETED PROTEIN OF AFRICAN SWINE FEVER VIRUS [J].
AFONSO, CL ;
ALCARAZ, C ;
BRUN, A ;
SUSSMAN, MD ;
ONISK, DV ;
ESCRIBANO, JM ;
ROCK, DL .
VIROLOGY, 1992, 189 (01) :368-373
[2]   HIGHLY SPECIFIC CONFIRMATORY WESTERN-BLOT TEST FOR AFRICAN SWINE FEVER VIRUS-ANTIBODY DETECTION USING THE RECOMBINANT VIRUS PROTEIN P54 [J].
ALCARAZ, C ;
RODRIGUEZ, F ;
OVIEDO, JM ;
EIRAS, A ;
DEDIEGO, M ;
ALONSO, C ;
ESCRIBANO, JM .
JOURNAL OF VIROLOGICAL METHODS, 1995, 52 (1-2) :111-119
[3]  
Alcaraz C, 1990, J Vet Diagn Invest, V2, P191
[4]   PERSISTENCE OF AFRICAN SWINE FEVER ANTIBODY REACTIVITY ON ELISA AND IMMUNOBLOTTING ASSAYS [J].
ARIAS, M ;
ESCRIBANO, JM ;
SANCHEZVIZCAINO, JM .
VETERINARY RECORD, 1993, 133 (08) :189-190
[5]   LABORATORY DIAGNOSIS AND DISEASE OCCURRENCE IN THE CURRENT AFRICAN SWINE FEVER ERADICATION PROGRAM IN SPAIN, 1989-1991 [J].
BECHNIELSEN, S ;
ARIAS, ML ;
PANADERO, J ;
ESCRIBANO, JM ;
GOMEZTEJEDOR, C ;
BONILLA, QP ;
SANCHEZVIZCAINO, JM .
PREVENTIVE VETERINARY MEDICINE, 1993, 17 (3-4) :225-234
[6]   GENETIC-VARIATION OF AFRICAN SWINE FEVER VIRUS - VARIABLE REGIONS NEAR THE ENDS OF THE VIRAL-DNA [J].
BLASCO, R ;
DELAVEGA, I ;
ALMAZAN, F ;
AGUERO, M ;
VINUELA, E .
VIROLOGY, 1989, 173 (01) :251-257
[7]   LONG-TERM PERSISTENT INFECTION OF SWINE MONOCYTES/MACROPHAGES WITH AFRICAN SWINE FEVER VIRUS [J].
CARRILLO, C ;
BORCA, MV ;
AFONSO, CL ;
ONISK, DV ;
ROCK, DL .
JOURNAL OF VIROLOGY, 1994, 68 (01) :580-583
[8]  
DETRAY DE, 1957, AM J VET RES, V18, P299
[9]  
ESCRIBANO JM, 1989, AM J VET RES, V50, P1118
[10]  
GomezPuertas P, 1996, J VIROL, V70, P5689