Hybridoma cell strain secreting monoclonal antibody against rice ragged stunt virus and use of monoclonal antibody

Abstract

The invention discloses a hybridoma cell strain secreting a monoclonal antibody against rice ragged stunt viruses (RRSV) and use of the monoclonal antibody. Purified RRSV coat protein is used as an antigen to immunize a BALB/C mouse, and thus a hybridoma cell strain 4H5 which can be passed down steadily and can secrete monoclonal antibody against RRSV is obtained, wherein the collection number ofthe hybridoma cell strain 4H5 is CGMCCNo.5536. The result of Westernblot detection indicates the monoclonal antibody secreted by the hybridoma cell strain 4H5 reacts with the RRSV coat protein specifically. The indirect enzyme-linked immuno sorbent assay(ELSA) titer of the hybridoma cell strain 4H5 monoclonal antibody ascitic fluid reaches 10<-7>, the type and subtype of the antibody is IgG1 and kappa chain. According to a dot-ELISA detection method established by using the 4H5 monoclonal antibody for detecting brown rice plant hopper and RRSV in rice, when single-head brown rice plant hopperis diluted to 800mu L and affected leaves is diluted according to a weight-volume (g/mL) ratio of 1:320, viruses can still be detected. The established dot-ELISA method can detect brown rice plant hopper and RRSV in rice accurately, specifically and sensitively. The preparation of the monoclonal antibody against RRSV and the establishment of the detection method thereof provide technical and material supports for diagnosing, predicting and forecasting and scientifically preventing and controlling rice virus diseases.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell strain secreting monoclonal antibody against rice tooth dwarf virus and the application of the monoclonal antibody. Background technique [0002] Rice ragged stunt virus (RRSV) is a new virus discovered in the late 1970s. It occurs locally in some countries in Southeast Asia, East Asia and South Asia, and has a certain impact on rice production. At present, it mainly occurs in Guangxi, Fujian, Hunan, Jiangxi, Guizhou, Yunnan and other places in China. The diseased plants are dark green and dwarfed, the leaves are twisted into jagged notches, and there are slender veins on the back of the leaves and the leaf sheaths. The onset before the tillering stage can not be heading, and the onset at the later stage will cause symptoms such as incomplete heading and insufficient grain. Infected fields generally reduce production by 10% to 20%, and serious production reductions...

AI Technical Summary

Problems solved by technology

In order to grasp the natural population of BPH and the disease status of rice, strengthen the early monitoring and early warning of rice leaf dwarf disease in my country, and scientifically guide the prevention and control, it is urgent to establish a high-throughput detection method for the detection of RRSV in BPH and rice, but there is currently no This high-throughput detection method can detect small samples only with low-efficiency electron microscope observation, RT-PCR method, nucleic acid electrophoresis, etc.

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