分泌抗甘薯羽状斑驳病毒单克隆抗体的杂交瘤细胞株构建及抗体初步应用

Construction of Hybridomas Secreting Monoclonal Antibodies Against Sweet Potato Feathery Mottle Virus and Use of Antibody for Detection of SPFMV

甘薯羽状斑驳病毒(Sweet Potato Feathery Mottle Virus,SPFMV),粒体条状,长800—850nm,不仅通过甘薯块根传播,而且被多种蚜虫非持久性传播,在世界甘薯产区均有发生,引起甘薯叶片明脉,脉带。褪绿斑及紫色羽状纹等症状,是我国甘薯产区的主要病毒之一。

Two hybridomas stably secreting monoclonal antibodies(MCAs) against sweet potato feathery mottle virus (SPFMV) were constructed by fusing mouse myeloma (Sp2/0-Ag14) with mouse spenocytes immunized with SPFMV-LF. MCAs secreted by the two hybridomas specifically reacted with both SPFMV——LF (China strain) and SPFMV——RC(American strain) in Dot blot-ELISA. A large amounts of MCA was produced with BALB/c mice. The titre of hybridoma cultured supernatant was 1:256, and that of ascitic fluids was 1:1×10~4. Immunoglobulins of MCAs were IgG2a and IgG3 respectively. The detection of SPFMV-infected sweet potato plants with MCAs confirmed that the MCAs obtained in this test were desirable for large scale routine surveys of SPFMV. The study on construction of hybridomas and detection of SPFMV using MCAs were the first report in the world.