Hybridoma cell strain secreting PPV (plum pox virus)-resistant monoclonal antibody and application of monoclonal antibody

Abstract

The invention discloses a hybridoma cell strain secreting a PPV (plum pox virus)-resistant monoclonal antibody and an application of the monoclonal antibody. Prokaryotic-expression PPV coat protein istaken as an antigen to immunize a BALB / c mouse, one hybridoma cell strain 4A8 capable of secreting the PPV-resistant monoclonal antibody is obtained by cell fusion, screening and cloning, and the preservation number of the hybridoma cell strain is CGMCC No.17283. Indirect ELISA valence of monoclonal antibody ascites secreted by the cell strain reaches 10<-7>, the type and the subclass of the antibody are IgG1 and kappa light chains, and the monoclonal antibody has a specific reaction with coat protein subunit of PPV. TAS-ELISA and dot-ELISA detection methods for detecting the PPV in plum trees are established on the basis of the monoclonal antibody 4A8, wherein sensitivity of the TAS-ELISA and dot-ELISA methods for detecting sick leaves reaches 20480-fold dilution and 5120-fold dilution (w / v,g / mL). The material and technical support is provided for diagnosis and detection of PPV disease in an orchard, import and export port inspection and quarantine as well as scientific prevention and control through establishment of the preparation and detection methods of the PPV monoclonal antibody.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell strain secreting anti-plum pox virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Plum pox virus disease was first discovered in Bulgaria in 1915, and then spread to the entire Europe and the United States, causing serious damage to stone fruit trees in the affected areas. The pathogen Plum pox virus (Plum pox virus, PPV) has been distributed in Europe, Africa, North America, South America and Asia since Atanasoff discovered it on plum trees. Since 2001, my country has successively discovered plum pox virus in Hunan, Nanjing, and Xi'an. The Ministry of Agriculture and Rural Affairs of my country has attached great importance to it and included plum pox virus in the "List of Imported Plant Quarantine Pests of the People's Republic of China". [0003] PPV is a virus of the genus Potyvirus belonging to the family Pot...

AI Technical Summary

Problems solved by technology

At present, there have been reports on the preparation of PPV antibodies and serological methods, but the detection sensitivity of the reported PPV antibodies and serological methods is not good, and the accuracy rate is not high

Images