Hybridoma cell strain capable of secreting tomato yellow leaf curl virus monoclonal antibody and application of monoclonal antibody

Abstract

The invention discloses a hybridoma cell strain capable of secreting a tomato yellow leaf curl virus (TYLCV) monoclonal antibody and application of the monoclonal antibody. A coat protein gene of a tomato yellow leaf curl virus separator (TYLCV-SH2) is cloned; the coat protein of the virus is expressed by a prokaryotic expression system; BALB/c mice are immunized by using expression and purification protein as antigen; a hybridoma cell strain D10 capable of performing passage stably and secreting the TYLCV monoclonal antibody is obtained through cell fusion, screening and cloning; and the collection number is CGMCC No.5538. The D10 monoclonal antibody ascites indirect ELISA valence reaches more than 10<-6>; and the antibody type and subclass are IgG1 and kappa chains. A dot-ELISA detection method for detecting the TYLCV of the tomatoes is established by the D10 monoclonal antibody; and when the leaf suffering from the disease is diluted according to the ratio of 1:320 (w/v and g/mL), the virus can still be detected. By the dot-ELISA and Tissue-blot ELISA method, the TYLCV of tomato samples in the field can be detected accurately, specifically and sensitively. Due to establishment of a preparation method for the TYLCV monoclonal antibody and a detection method, technological and material support is provided for diagnosis, prediction and scientific prevention and control of the tomato virus disease.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell strain secreting anti-tomato yellow leaf curl virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Tomato is an important vegetable crop in the world. It has many varieties, wide adaptability and rich nutrition. It is one of the 30 crops with the highest total output in the world. The harm of geminiviruses to tomato is one of the important restrictive factors of tomato production, especially in recent decades, the spread of geminiviruses worldwide has caused its harm to tomato is becoming more and more serious. Among all the geminiviruses infecting tomato, tomato yellow leaf curl virus (Tomato yellow leaf curl virus, TYLCV) is the most serious. TYLCV was first discovered and officially named in Israel in 1964. In the following 30 years, the disease spread rapidly to the Middle East, Mediterranean coast, Southeast As...

AI Technical Summary

Problems solved by technology

In addition, the climatic conditions, varieties and cultivation methods in different regions are different, and it is difficult to achieve effective control effect by one method alone. Therefore, comprehensive control measures adapted to local conditions should be adopted for the control of TYLCV, and a set of sensitive and rapid virus control measures should be established for this purpose. The detection method is of great significance for the monitoring and prevention of the virus

At present, only low-efficiency electron microscope observation, RT-PCR method and nucleic acid hybridization are used to detect viruses in small samples.

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