Hybridoma cell strain excreting monoclonal antibody (MAb) resisting rice blackstreaked dwarf virus (RBSDV) and application of MAb

Abstract

The invention discloses a hybridoma cell strain excreting a monoclonal antibody (MAb) resisting a rice blackstreaked dwarf virus (RBSDV) and the application of the MAb. A coat protein (CP) of the RBSDV is expressed into antigen immune BALB / c mouse by a prokaryotic expression method, and the hybridoma cell strain 5G1 which can stably passage and excrete the MAb resisting the RBSDV is obtained through cell fusion, selection and cloning, wherein the preserving number is CGMCC No. 5537. Ascitic indirect enzyme-linked immuno sorbent assay (ELISA) valence of the 5G1 MAb is over 10<-6>, and the antibody type and the subtype are IgG1 and kappa chain. The antibody excreted by the cell strain and the coat protein of the RBSDV have specific immunobinding reaction. By the dot-ELISA detection method for detecting rice planthopper and the RBSDV in rice and established by using the 5G1 MAb as a core, the virus can be still detected when a single-head small brown rice planthopper is diluted to 1,600 microlitres and sick leaves are diluted (w / v, g / mL) in the proportion of 1:160. The preparation of the MAb resisting the RBSDV and the establishment of the detection method thereof provide technical and material support for the diagnosis, forecast and scientific prevention and control of rice viruses.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell line secreting a monoclonal antibody against rice black-streaked dwarf virus and the application of the monoclonal antibody. Background technique [0002] In the mid-1960s, rice black-streaked dwarf disease caused severe damage to cereal crops such as rice, wheat, and corn in many provinces and cities in Zhejiang and East China. The corn planting area was forced to change the cultivation system, and the diseased area decreased rapidly in the next 20 years until it disappeared in the 1970s. In recent years, black-streaked dwarf disease has occurred and prevailed in Zhejiang, Shanghai, Jiangsu, Anhui, Jiangxi and other places due to various factors such as changes in cultivation system, climate warming, and outbreaks of SBPH. [0003] Rice black streaked dwarf disease is caused by rice black streaked dwarf virus (RBSDV), which belongs to the genus Fijivirus of the fam...

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Problems solved by technology

In order to grasp the natural population of SBPH and the disease status of rice, strengthen the early monitoring and early warning of rice black-streaked 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 RBSDV in SBPH and rice , but currently there is no such high-throughput detection method, only low-efficiency electron microscope observation, RT-PCR method and other methods are used for small sample detection

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