Hybridoma cell strain secreting citrus yellow vein clearing virus-resistant monoclonal antibodies and monoclonal antibody application thereof

Abstract

The invention discloses a hybridoma cell strain secreting citrus yellow vein clearing viru(CYVCV)-resistant monoclonal antibodies and monoclonal antibody application thereof. Coat protein of prokaryotic-expression CYVCV serves as antigens to immune BALB/c mice, cell fusion, screening and cloning are performed, one hybridoma cell strain 18H5 which is stable in passage and can secrete CYVCV-resistant monoclonal antibodies is obtained, and the preservation number of the hybridoma cell strain 18H5 is CGMCC No. 12003. The ascites indirect ELISA titer of the monoclonal antibodies secreted by the cell strain reaches 10<-7>, according to the antibody type and the subclass, the monoclonal antibodies are composed of IgG1 and kappa light chains, and the monoclonal antibodies and CYVCV coat protein subunits of 32 kD have a specific reaction. The 18H5 monoclonal antibodies are used for establishing TAS-ELISA, dot-ELISA and Tissue blot-ELISA detecting methods of CYVCV in citrus trees, wherein the diseased leaf detecting sensitivity of the TAS-ELISA method and the diseased leaf detecting sensitivity of the dot-ELISA method reach 1:2560 and 1:20480 fold dilution (w/v, g/mL) respectively. Preparation of the CYVCV-resistant monoclonal antibodies and establishment of the detection methods of the CYVCV-resistant monoclonal antibodies provide matter and technologic support for detection and diagnosis, epidemiological analysis and scientific prevention and control of citrus virus diseases.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell line secreting monoclonal antibody against citrus yellowing vein brightening virus and the application of the monoclonal antibody. Background technique [0002] Citrus yellow vein clearing disease is caused by Citrus yellow vein clearing virus (CYVCV), a member of the genus Citrus virus in the Alphaflexiviridae family. The disease was first discovered in Pakistan in 1988, and is currently spreading rapidly in citrus producing areas in Pakistan, India, Turkey and other countries, causing serious losses to the citrus industry. The disease causes yellowing and bright veins of the leaves of citrus plants such as lemons and limes, accompanied by symptoms such as leaf curling and shrinkage. In severe cases, the leaf veins can be necrotic and the young leaves will fall off, resulting in weak tree vigor and reduced yield. . Recently, citrus trees infected with CYVCV were s...

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Problems solved by technology

At present, the detection and diagnosis of CYVCV only use low-efficiency electron microscope obse

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