Submitted to: Revista Mexicana de Fitopatologia
Publication Type: Peer reviewed journal
Publication Acceptance Date: 11/29/2006
Publication Date: 12/1/2006
Citation: Sandoval-Alejo, B.D., Iracheta-Cardenas, M.M., Keremane, M.L., Lee, R.F., Rocha-Pena, M.A. 2006. Evaluación de Anticuerpos Desarrollados Contra la Proteína Recombinante de la Cápside del Virus Tristeza de los Cítricos. Revista Mexicana de Fitopatologia, Vol. 24: 137-144. Interpretive Summary: Development of a reliable source of antibodies which are available in enough quantity for use in large scale assays is a continuing problem for the diagnosis of several viruses which are hard to purify. This research reports on cooperative research for the production of antibodies specific for Citrus tristeza virus (CTV) by injecting rabbits and goats with the expressed coat protein gene of three isolates of CTV. The effectiveness of the resultant antibodies for detection of CTV in isolates from the CTV isolate collection in Mexico is demonstrated.
Technical Abstract: Polyclonal antibodies specific for the recombinant coat protein (rCP) p25 gene of (Citrus tristeza virus = CTV), were developed for isolates MX08 and MX14 from México and B227 from India. The reactivity of rCP antibodies was evaluated using healthy and CTV infected tissue. The combination of rCP antibodies developed in goats for isolate MX14 used as primary (trapping) antibodies, and rCP antibodies developed in rabbits for isolates MX08, MX14 and B227 as for antigen detection, reacted efficiently, with optical density (OD405) values of between 0.400 to 1.400 for CTV infected tissue and less than 0.100 for healthy tissue. The combination of rCP antibodies T1(goat anti-CTV MX14) as primary antibody and C3 (rabbit anti-CTV MX08) as detection antibody was as equally effective as the commercial kit from Agdia for positive and negative sample discrimination from a collection of 32 CTV isolates from Nuevo Leon, Mexico. In addition, the rCP from isolates MX14 and B227 mixed with healthy tissue, were equally effective as positive control in ELISA tests as extracts fro CTV infected plants, with OD405 values in the range of 0.300 and 0.400.