Hybridoma cell strain secreting monoclonal antibody against citrus tatter leaf virus and application of monoclonal antibody thereof

Abstract

The invention discloses a hybridoma cell strain secreting a monoclonal antibody against citrus tatter leaf virus, and an application of the monoclonal antibody thereof. A coat protein gene of citrus tatter leaf virus (CTLV) is cloned, and is expressed by a prokaryotic expression system; a BALB / c mouse is immunized with the expression-purified protein as an antigen, and a hybridoma cell strain 6C5 which can be subcultured stably and secretes a monoclonal antibody against CTLV is obtained through cell fusion, screening, and cloning, and the accession number is CGMCC No. 8779. The monoclonal antibody has an ascetic indirect ELISA titer of up to 10<-7>, and both the antibody type and subtype are IgG1, kappa chain. A dot-ELISA method for detecting CTLV in citrus established with the monoclonal antibody can detect the virus even when a diseased leaf is diluted by 1:640 times (w / v, g / mL). The acquisition of the hybridoma cell secreting monoclonal antibody against CTLV and its monoclonal antibody, and the establishment of a related serological detection method provide technical and material support for the diagnosis, prediction and scientific prevention and control of citrus viral diseases.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell strain secreting anti-citrus leaf leaf virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Citrus tatter leaf virus (CTLV) caused by citrus tatter leaf virus (CTLV) is widely distributed in my country, including Taiwan, Zhejiang, Guangxi, Hunan, Hubei and Sichuan provinces (regions). An important citrus viral disease following split skin disease. The citrus varieties Wenzhou Tangerine and Beijing Lemon that originated in my country are considered to be the original source of the virus. Because the susceptible scion of Beijing Lemon is on the thick-skinned Lemon plant, it can cause yellow spots on the leaves, become smaller, and the leaf edges are broken, so it is called Leaf breakage. The occurrence of CTLV is also very common in the adjacent areas of my country such as South Korea, Japan, and the Philippines, and C...

AI Technical Summary

Problems solved by technology

In order to control the incidence of citrus, strengthen the early monitoring and early warning of citrus leaf fragment 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 CTLV in citrus. At present, there is no such high-throughput detection method. Low-efficiency electron microscope observation, RT-PCR method, nucleic acid electrophoresis and other methods for small sample detection

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