Hybridoma cell line secreting monoclonal antibody against rice tumor dwarf virus and application of monoclonal antibody

Abstract

The invention discloses a use of a hybridoma cell line secreting a monoclonal antibody against rice gall dwarf viruses and a use thereof. A BALB / c mouse is immunized by a prokaryotically expressed rice gall dwarf virus (RGDV) CP recombinant protein as an antigen, and a hybridoma cell line 11B6 capable of stable passage and secretion of an anti-RGDV monoclonal antibody is obtained by cell fusion, screening and cloning and has a preservation number of CGMCC No. 14897. The indirect ELISA titer of the ascites monoclonal antibody secreted by the hybridoma cell line is 10<-8>. The antibody type andsubclass are IgG1 and kappa chains. The monoclonal antibody can react specifically with the RGDV coat protein. The ACP-ELISA and dot-ELISA methods for detecting RGDV are established through a 11B6 monoclonal antibody as the core. The sensitivity of the two methods for detecting diseased leaves is 1:327 680-fold dilution (w / v, g / mL) and 1:20 480-fold dilution (w / v, g / mL). The acquisition of the hybridoma cell line and its monoclonal antibody and the establishment of serological detection methods provide material and technical support for the detection, diagnosis and scientific prevention and control of the viral disease.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell line secreting anti-rice tumor dwarf virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Rice gall dwarf disease is a viral disease of rice caused by rice gall dwarf virus (RGDV). Since it was discovered in Thailand in 1979, rice tumor dwarf disease has been found in Malaysia, Japan, South Korea and other places one after another. RGDV was first discovered in Guangdong, my country in 1981. At present, rice tumor dwarf disease is mainly prevalent in southern rice areas such as Guangdong, Guangxi, Fujian, and Hainan. Fields with severe disease may not even be able to head normally, and the grains will not be harvested. Besides rice, the host plants of RGDV also include gramineous crops such as wheat, barley, oats, apricot, wild rice and corn, but it does not infect barnyardgrass. Typical symptoms of RGDV-infected ri...

AI Technical Summary

Problems solved by technology

At present, judging the occurrence of rice tumor dwarf virus in the field based on symptom observation is still the most important method, but rice has no symptoms in the early stage of virus infection, and the symptoms caused by various viral diseases are very similar. In addition, there are often cases of virus compound infection , so the results obtained by relying on symptom observation lack accuracy and scientific

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

Although the molecular detection technology based on the PCR method is sensitive, it is not suitable for the detection of large-scale samples in the field

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