Hybridoma cell line secreting anti-wheat dwarf virus monoclonal antibody and its application

Abstract

The invention discloses a hybridoma cell strain capable of secreting a wheat dwarf virus resisting monoclonal antibody and a monoclonal antibody application of the strain. WDV (wheat dwarf virus) CP recombinant protein in prokaryotic expression is used as an antigen immune for BALB / c mice, the hybridoma cell strain 23F4 capable of stably going down to the future generation and secreting the WDV antibody is obtained after cell fusion, screening and cloning, and the collection number of the strain is CGMCC No. 13297. The abdominal dropsy indirect ELISA (enzyme-linked immuno sorbent assay) valence of the antibody secreted by the hybridoma cell strain reaches 10<-9>, and the antibody type and subclass are IgG1 and kappa chains. The antibody and WDV coat protein have specific reaction. The 23F4 antibody is used as a core for building ACP-ELISA and dot-ELISA methods for detecting WDV in a wheat plant, and diseased leaf detection sensitivity of the two methods reach 1:163840 and 1:5120 times of dilution (w / v, g / mL) respectively. Acquisition of the hybridoma cell strain and the antibody thereof and building of serological detection methods provide substantial and technical support for virus disease detection, diagnosis 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 anti-wheat dwarf virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Wheat dwarf disease (Wheat dwarf disease) is one of the main viral diseases that endanger wheat production. In 1961, Vacke discovered the dwarf phenomenon of wheat in Czechoslovakia. He believed that it was caused by a virus transmitted by leafhoppers, and named it Wheat dwarf virus (WDV). In China, wheat dwarf disease was first discovered in Hancheng, Shaanxi in 2007. At that time, the diseased area reached 10,000 mu, and the average rate of diseased plants in the field reached 80%. At present, the disease has also occurred in 13 major wheat-producing provinces in my country, including Hebei, Yunnan, Shanxi, Gansu, and Henan. [0003] The diseased plants of wheat dwarf disease often appear in lumps in the field, and its typical ...

AI Technical Summary

Problems solved by technology

At present, judging the occurrence of wheat dwarf disease in the field based on symptom observation is still the most important method, but because the symptoms caused by various viral diseases are very similar, and are often co-infected with other viral diseases, the results obtained only by symptom observation lack of accuracy

Electron microscope observation of virus particles requires expensive instruments, and the size and shape of virus particles of different viral diseases are similar, so it is only used as an auxiliary detection method

Although the molecular detection technology based on PCR method is sensitive, it is not suitable for field detection

Relatively speaking, serological methods are simple, easy to operate, fast, sensitive, and low in cost, and can be applied to large-scale detection of field samples. of high-quality antibodies

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