Improved serological diagnosis of Poplar mosaic virus with monoclonal antibodies.
Poplar mosaic virus (PopMV) is widespread in all countries where poplar is grown, and causes severe economic losses in terms of quantity and quality of wood production. Control is based on indexing, aimed at the production of healthy propagation material. The currently used diagnostic method is double antibody sandwich (DAS) ELISA with polyclonal antibodies, which is relatively simple and inexpensive and more reliable than visual inspection of symptoms in the nurseries. However, this method also has disadvantages, mainly low sensitivity in relation to low concentration and irregular distribution of the virus in the plant. In this study, a new diagnostic method for PopMV based on production and use of a monoclonal antibody (Mab) in a triple antibody sandwich (TAS) ELISA, is presented. The TAS-ELISA with monoclonal antibodies was optimised by testing a range of reagent combinations and concentrations. PopMV was detected by the optimised TAS-ELISA with sensitivity more than 100 times higher than by DAS-ELISA with polyclonal antibodies. Six PopMV isolates from four European countries were detected with the same efficiency, indicating that no limitations to the practical use of the TAS-ELISA arise due to excessive epitope-specificity of the monoclonal antibody employed.
Carra
A
A
Dipartimento di Colture Arboree, Università di Torino, via L. da Vinci 44, 10095 Grugliasco (TO), Italy. andrea.carra@unito.it
Brocchi
E
E
Simone
F De
FD
Luisoni
E
E
D000818:Animals; D000911:Antibodies, Monoclonal; D001042:Aphids / Q000821:virology; D017788:Carlavirus / Q000145:classification / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D007106:Immune Sera; D010935:Plant Diseases / Q000821:virology*; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D007106:Immune Sera