Detection of Citrus tristeza virus with Antibodies Developed to the Recombinant p25 Capsid Protein by Direct Tissue Blot Immunoassay

摘要

Citrus tristeza virus (CTV) is widespread and causes one of the most economical important viral diseases of citrus. The large scale analysis of citrus samples for CTV detection is conducted routinely by the enzyme-linked immunosorbent assay (ELISA) orits variant named direct tissue blot immunoassay (= immunoprinting). In this study, we evaluated the use of polyclonal antibodies developed to recombinant p25 CTV coat protein for virus detection under an immunoprinting system by using healthy and fieldCTV infected citrus samples. The combination of goat (CB) anti-CTV/IgG anti-goat-AP conjugate provided a positive reaction with CTV infected plants, and negative with healthy citrus. In addition, the combination of goat CB anti-CTV coupled with biotin,plus avidin-AP as commercial conjugate was equally effective in discriminating CTV infected from healthy citrus. Furthermore, antibodies developed to the recombinant p25 CTV coat protein, either untagged or biotin conjugated showed comparable results with commercially available immunoprinting kits for CTV detection by Plant Print and Agdia.