Hybridoma cell line secreting monoclonal antibody against citrus chlorosis virus and application of monoclonal antibody

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 present invention relates to the field of biotechnology, in particular to a hybridoma cell strain that secretes monoclonal antibodies against Citrus Yellow Venom Virus and the application of the monoclonal antibodies thereof. Background technique [0002] Citrus yellow vein clearing virus is caused by Citrus yellow vein clearing virus (CYVCV), which belongs to the Alphaflexiviridae family Citrus virus. 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 of the veins and clear veins of the leaves of citrus plants such as lemons and limes, and is accompanied by leaf rewinding and shrinking. In severe cases, it can lead to necrosis of leaf veins and shedding of tender leaves, resulting in weak tree vigor and decreased yield. . Recently, citrus trees infected with CYVCV ha...

AI Technical Summary

Problems solved by technology

At present, the detection and diagnosis of CYVCV only use low-efficiency electron microscope observation, RT-PCR method, nucleic acid electrophoresis and other methods for small sample detection

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