Generation of specific antibody against recombinant major coat protein of Beet necrotic yellow vein virus and its application for detection of rhizomania disease

Abstract

Beet necrotic yellow vein virus (BNYVV) is one of the main contributors to economic losses in sugar beet production. The present study generated a polyclonal antibody that detects BNYVV. The conserved genomic region of all BNYVV isolates gene encoding the viral major coat protein (BNYVV-CP) was amplified, cloned, sequenced, and expressed in E. coli strain BL21 (DE3). The expressed protein was purified under native conditions by affinity chromatography. The purified BNYVV CP was used as a recombinant antigen to raise polyclonal antibodies in rabbits. Purified anti-CP-IgG detected the BNYVV-CP recombinant protein and BNYVV in infected sugar beet by indirect-ELISA, dot immunobinding assay, and western blot analysis blotting. The serological assays strongly proved the sensitivity and specificity of anti-BNYVV-CP-IgG against BNYVV. These results suggest that the generated anti-BNYVV-CP-IgG polyclonal antibodies can be used as reliable and quick means for the BNYVV virus detection under field conditions.