Hybridoma cell line secreting monoclonal antibody against phoenix mosaic virus and its application

Abstract

The invention discloses a hybridoma cell strain capable of secreting an anti-Pepino mosaic virus (PepMV) monoclonal antibody, and an application of the monoclonal antibody. Purified PepMV virus particles are used as an antigen to immunize BALB / c mice, the hybridoma cell strain 23D11 capable of secreting the anti-PepMV monoclonal antibody is obtained through a hybridoma technology, and the preservation number of the hybridoma cell strain 23D11 is CGMCC No.18845. The ascites indirect ELISA titer of the monoclonal antibody secreted by the cell strain reaches 10<-8>, the antibody type and subclassare IgG1 and kappa light chain, and the monoclonal antibody has specific immune reaction with PepMV, does not react with potato virus X belonging to the same genus, and does not have immune reactionwith tomato mottle mosaic virus, tomato mosaic virus, tomato spotted wilt virus, tomato yellow leaf curl virus, tomato black loop virus and healthy plant tissues. A variety of detection methods are established by using the 23D11 monoclonal antibody, and the sensitivity of ACP-ELISA and dot-ELISA methods for detecting diseased leaves reaches 1:40, 960 fold dilution and 1:20, 480 fold dilution (w / v,g / mL) respectively. The creation of the cell strain and the establishment of a virus serological detection method provide material and technical support for PepMV detection and diagnosis, epidemiological investigation and scientific prevention and control.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell strain secreting monoclonal antibody against phoenix fruit mosaic virus and the application of the monoclonal antibody. Background technique [0002] Pepino mosaic virus (PepMV) is a member of the Potexvirus genus of the family Flexiviridae, and its genome is positive-sense single-stranded RNA. The genome of PepMV contains five open reading frames, expressing five proteins respectively. The first reading frame encodes a 164kDa RNA polymerase, and the middle three reading frames encode 26, 14, and 9kDa proteins to form a three-gene cassette, which is involved in the virus Cell-to-cell movement, the last reading frame encodes a 27kDa coat protein. First identified on Dutch tomatoes in 2000, the virus is now a major viral disease of greenhouse tomato crops worldwide. [0003] PepMV can infect a variety of Solanaceae plants, especially causing serious damage to tomato....

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Problems solved by technology

However, the establishment of serological methods depends on specific high-quality virus antibodies

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