Hybridoma cell strain secreting monoclonal antibody resistant to tomato black ring nepovirus and application of monoclonal antibody of hybridoma cell strain

Abstract

The invention discloses a hybridoma cell strain secreting a monoclonal antibody resistant to tomato black ring nepovirus and application of the monoclonal antibody of the hybridoma cell strain.Purified tomato black ring nepovirus (TBRV) particles serve as antigen to immunize a BALB / c mouse, cell fusion, screening and cloning are conducted, the hybridoma cell strain 12H3 which can stably subculture and secrete the TBRV-resistant monoclonal antibody is obtained, and the preservation No.of the hybridoma cell strain is CGMCC No.12005.The titer of ascites indirect ELISA of the monoclonal antibody secreted by the hybridoma cell strain reaches 10<-7>, and the type and subclass of the antibody is an IgG1,kappa light chain.The monoclonal antibody has a specific immunoreaction with 50kDa TBRV coat protein.An ACP-ELISA method and a dot-ELISA method for detecting TBRV in tomatoes are established through the 12H3 monoclonal antibody, and sensitivity of the ACP-ELISA method and sensitivity of the dot-ELISA method for detecting leaves attacked by disease reach 1: 163, 840 and 1: 5, 120 times dilution (w / v, g / mL) respectively.Through obtaining of the hybridoma cell strain 12H3 and the monoclonal antibody and establishment of the serological test methods, material support and technical support are provided for detection, quanrantine and scientific prevention and control for the virus.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell line secreting anti-tomato black ring virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Tomatoblackringnepovirus (TBRV) belongs to the genus Nepovirus of the family Comoviridae, and was first discovered by Smith on tomatoes in the UK in 1946, followed by Italy, Germany, Romania, Hungary, France Researchers in China, Poland, the United Kingdom and other places have also isolated tomato black ring virus from various plants. It can be spread over long distances both by seeds and through soil and plant propagation material. Therefore, TBRV has become a global crop virus, which is distributed in many European countries such as France, Finland, Sweden, Russia, Germany, and also in some Asian countries such as India, Japan and Turkey, the United States, Canada, Kenya It has also been found in Brazil and Brazil, and it ...

AI Technical Summary

Problems solved by technology

At present, there have been many reports on the molecular biology technology of tomato black ring virus inspection and quarantine at home and abroad, but there are few reports on the application of serological detection. Although the molecular biology method is specific and sensitive, it requires expensive and sophisticated instruments and is difficult to operate. High technical requirements for personnel, not suitable for mass sample testing

The serological method is simple to operate, has good specificity, and can detect a large number of samples on a large scale, but it must rely on specific, sensitive and high-quality antibodies

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