Hybridoma cell strain capable of secreting BYDV (barley yellow dwarf virus) PAV strain monoclonal antibody and application of monoclonal antibody of hybridoma cell strain

Abstract

The invention discloses a hybridoma cell strain capable of secreting a BYDV (barley yellow dwarf virus) PAV strain monoclonal antibody and an application of the monoclonal antibody of the hybridoma cell strain. Purified BYDV PAV strain virions are used as an antigen to immunize a BALB / c mouse, one hybridoma cell strain 24G4 capable of passaging steadily and secreting the BYDV PAV strain monoclonal antibody is obtained through cell fusion, screening and cloning, and the preservation number of the hybridoma cell strain 24G4 is CGMCC No.13298. The indirect ELISA titer of monoclonal antibody ascites secreted by the hybridoma cell strain is 10<-9>, and the antibody type and subtype are IgG1 and kappa chain. ACP-ELISA and dot-ELISA methods for detecting BYDV PAV in wheat plants are established with the monoclonal antibody 24G4 as the core, and the sensitivities of the two methods for detecting infected leaves reach 1:81920 and 1:1280 fold dilution (w / v, g / mL). The hybridoma cell strain 24G4, the monoclonal antibody obtained from the same and the established serological test methods provide material and technical support for detection, diagnosis and scientific prevention and control of the viral diseases.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell line secreting monoclonal antibody against barley yellow dwarf virus PAV strain and the application of the monoclonal antibody. Background technique [0002] Wheat yellow dwarf disease caused by Barley yellow dwarf viruses (BYDVs) is one of the important viral diseases of wheat. The disease was first discovered by Oswald and Houston in 1951 on barley in California, USA. Currently, wheat yellow dwarf virus is distributed all over the world. In 1960, BYDVs appeared for the first time in Northwest China, and the yield of wheat infected by BYDVs generally decreased by 40%, and in severe cases, it could reach more than 70%. Therefore, BYDVs is also called "wheat cancer" and "yellow plague". Since it was first discovered, wheat yellow dwarf disease broke out several times on a large scale in the 1960s to 1980s, posing a major threat to my country's wheat production and f...

AI Technical Summary

Problems solved by technology

[0006] At present, the field often judges the occurrence of wheat yellow dwarf disease according to the observation of symptoms, but compared with fungal diseases and bacterial diseases, the symptoms of viral diseases are not obvious, and are often confused with other viral diseases, resulting in misjudgments, and through symptoms It is not possible to determine the specific strain of the virus

Electron microscope observation requires expensive equipment and professionals, and is not suitable for grassroots applications

Although molecular biology methods such as RT-PCR have high sensitivity, they are often limited by instruments and are only suitable for the detection of small samples in the laboratory

In comparison, serological methods are simple, easy to operate, fast, sensitive, and low in cost. They are currently ideal detection methods for plant viruses, but serological methods must rely on specific, sensitive and high-quality antibodies

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