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.
Oviedo
J M
JM
Centro de Investigación en Sanidad Animal (CISA-INIA), Madrid, Spain.
Rodríguez
F
F
Gómez-Puertas
P
P
Brun
A
A
Gómez
N
N
Alonso
C
C
Escribano
J M
JM
D000357:African Swine Fever / Q000175:diagnosis / Q000276:immunology / Q000821:virology*; D000358:African Swine Fever Virus / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D016367:Baculoviridae; D015153:Blotting, Western; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D010750:Phosphoproteins / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D018411:Spodoptera / Q000166:cytology; D013552:Swine; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics* / Q000276:immunology; D015678:Viral Structural Proteins / Q000235:genetics* / Q000276:immunology
D000914:Antibodies, Viral; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C093921:p54 protein, African swine fever virus; C074960:P30 protein, African Swine Fever Virus