Pepino mosaic virus （PepMV）, monopartite RNA virus, 6,500 pb, belonging to Flexiviridae and Potexvirus group, is highly infectious and easily transmissible. Its economic impact is major for the tomato producer＇s co...Pepino mosaic virus （PepMV）, monopartite RNA virus, 6,500 pb, belonging to Flexiviridae and Potexvirus group, is highly infectious and easily transmissible. Its economic impact is major for the tomato producer＇s countries. Prevention, based on early virus detection is the only effective control measure. Monoclonal antibodies appeared to be very useful tool. The authors used for the production of monoclonal antibodies hybridomas technique, by fusing spleen cells of immunized BALB/c mice to PepMV and SP2/O cancerous cells. The aim of this work is to produce hybridomas producers of Mab that could be used for ELISA in Morocco. At the same time, these efforts will serve to decrease expenses of producers concerning phytosanitory control. We obtained 16 hybridomas lines producers of Mab specific for PepMV. They were tested for efficiencies in ELISA and two lines were retained for production of Mab on large scale （1B 11-G 10 and 5A l-G5）. Isotyping of these two lines showed that they are belonging to IgG 1 class and easily purified by affinity chromatography in agarose column by protein A. The conjugation of these two antibodies to alkaline phosphatase has been verified by DAS-ELISA. These antibodies will enable to diagnose the disease from infected tomato plants, integrating several serological tests to control it and target the actions of struggles.展开更多
文摘Pepino mosaic virus （PepMV）, monopartite RNA virus, 6,500 pb, belonging to Flexiviridae and Potexvirus group, is highly infectious and easily transmissible. Its economic impact is major for the tomato producer＇s countries. Prevention, based on early virus detection is the only effective control measure. Monoclonal antibodies appeared to be very useful tool. The authors used for the production of monoclonal antibodies hybridomas technique, by fusing spleen cells of immunized BALB/c mice to PepMV and SP2/O cancerous cells. The aim of this work is to produce hybridomas producers of Mab that could be used for ELISA in Morocco. At the same time, these efforts will serve to decrease expenses of producers concerning phytosanitory control. We obtained 16 hybridomas lines producers of Mab specific for PepMV. They were tested for efficiencies in ELISA and two lines were retained for production of Mab on large scale （1B 11-G 10 and 5A l-G5）. Isotyping of these two lines showed that they are belonging to IgG 1 class and easily purified by affinity chromatography in agarose column by protein A. The conjugation of these two antibodies to alkaline phosphatase has been verified by DAS-ELISA. These antibodies will enable to diagnose the disease from infected tomato plants, integrating several serological tests to control it and target the actions of struggles.
