Monoclonal antibody hybridoma cell line for secreting southern rice black-streaked dwarf resistant viruses and monoclonal antibody application thereof

Abstract

The invention discloses a monoclonal antibody hybridoma cell line for secreting southern rice black-streaked dwarf resistant viruses and monoclonal antibody application thereof. Polypeptide of 12 amino acids at the C end of a capsid protein of southern rice black-streaked dwarf viruses (SRBSDVs) coupled with bovine serum albumin is used as an antigen to immune a BALB / c mouse, one hybridoma cell line 3F1 capable of performing stable passage and secreting an SRBSDV monoclonal antibody is obtained through cell fusion, screening and cloning, and the preservation number of the hybridoma cell line 3F1 is China general microbiological culture collection center number (CGMCCNo.) 5535. Monoclonal antibody ascites indirect enzyme-linked immunosorbent assay (ELISA) valence of the 3F1 reaches above 10-6, an antibody type and a subclass are immunoglobulin G (IgG1) and a kappa chain, the monoclonal antibody of the hybridoma cell line has specific reaction with a capsid protein subunit of SRBSDV56kD, and a method for detecting rice planthoppers and dot-ELISA of SRBSDV in rice is built by using the monoclonal antibody of the 3F1. When a single-head white backed rice planthopper is diluted to 6400mul, the viruses can still be detected when diseased leaves are diluted according to 1:320 times (w / v, g / mL). The preparation of the SRBSDV resistant monoclonal antibody and the building of the detection method provide technology and material supports for diagnosis, prediction and scientific prevention and control of the rice viruses.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell strain secreting anti-Southern rice black-streaked dwarf virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Southern rice black-streaked dwarf disease was first discovered in Yangxi County, Guangdong Province in 2001. Since then, the scope of occurrence has gradually expanded, and the damage has become increasingly serious. It spread to the Yangtze River Basin in 2008. In 2009, the viral disease occurred in an area of ​​about 400,000 hm in my country. 2 , the late rice was severely damaged, more than 6,700 hm 2 Rice harvest is basically lost. In 2010, the disease broke out in 13 southern provinces including Guangxi, Guangdong, Jiangxi, Hunan, Fujian, Guizhou, Yunnan, Zhejiang, Anhui, and Hainan, with an area of ​​about 1.33 million ha 2 , There are many spot failures in the middle and late rice areas, which hav...

AI Technical Summary

Problems solved by technology

In 2010, the disease broke out in 13 southern provinces including Guangxi, Guangdong, Jiangxi, Hunan, Fujian, Guizhou, Yunnan, Zhejiang, Anhui, and Hainan, with an area of ​​about 1.33 million hm2. There are many crop failures in the late rice area, causing huge losses to my country's rice production

In order to grasp the natural population of white-backed planthopper and the disease status of rice, strengthen the early monitoring and early warning of rice black-streaked dwarf disease in southern my country, and scientifically guide the prevention and control, it is urgent to establish a high-throughput detection of SRBSDV in white-backed planthopper and rice. At present, there is no such high-throughput detection method, and only low-efficiency electron microscope observation, RT-PCR method, nucleic acid electrophoresis and other methods are used for small sample detection

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