Differential diagnosis of Brazilian strains of Citrus tristeza virus by epitope mapping of coat protein using monoclonal antibodies.
Citrus tristeza virus (CTV) is one of the most important citrus pathogen, and among Brazilian CTV strains, the genotype Capão Bonito (CB) is the most harmful. Therefore, the coat protein (CP) gene were cloned and expressed as recombinant protein and used to develop four specific monoclonal antibodies (MAbs). Our previously data had showed these MAbs could recognize different strains of CTV and the present goal is to identify the epitopes of the recombinant CP by ELISA screening of overlapping recombinant peptides and to determine the binding specificity of CTV isolates in light of their antigenic domains onto CB strains. Three MAbs, 30.G.02, 37.G.11 and 39.07 recognized linear and no identical epitopes, but the fourth MAb, IC.04-12, probably had a conformational epitope, since it could not be identified by ELISA screening. Our previous data revealed MAb IC.04-12 do not recognize CP under denaturing conditions, but can identify weak CTV strains in ELISA involving crop samples. MAb 30.G.02 recognized an extremely conserved sequence and can be classified as "universal" antibody, and, interestingly, the epitope turned out by MAb 39.07 corresponded to severe CTV isolates. So, these MAbs can be applied in a differential screening by ELISA.
Peroni
Luís Antonio
LA
Departamento de Anatomia, Biologia Celular, Fisiologia e Biofísica, UNICAMP, Campinas, SP, Brazil.
Lorencini
Márcio
M
dos Reis
José Raimundo Ribeiro
JR
Machado
Marcos Antonio
MA
Stach-Machado
Dagmar Ruth
DR
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001938:Brazil; D036022:Capsid Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D002957:Citrus / Q000821:virology*; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D018604:Epitope Mapping; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000821:virology*; D010942:Plant Viruses / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D036022:Capsid Proteins; D004279:DNA, Viral; D011994:Recombinant Proteins