Hybridoma cell line secreting tomato mottle mosaic virus monoclonal antibody and its application

Abstract

The invention discloses a hybridoma cell line secreting anti-tomato mottle mosaic virus (ToMMV) monoclonal antibody and the application of the monoclonal antibody. BALB / c mice were immunized with purified ToMMV virus particles as antigen, and a hybridoma cell line 2D6, which can secrete anti-ToMMV monoclonal antibody, was obtained through cell fusion, screening and cloning, and its preservation number was CGMMC NO.18846. The cell line secretes monoclonal antibody ascitic fluid indirect ELISA titer up to 10 ‑8 , which has a specific immune reaction with ToMMV, but not with the same genus of tobacco mosaic virus (Tobacco mosaic virus, TMV) and tomato mosaic virus (Tomato mosaic virus, ToMV). ACP‑ELISA, dot‑ELISA and Tissue print‑ELISA detection methods for detecting ToMMV in tomato plants were established using 2D6 monoclonal antibody, among which the sensitivity of ACP‑ELISA and dot‑ELISA in detecting diseased leaves reached 1:40,960 and 1:20,480 times, respectively Dilution (w / v, g / mL). The creation of ToMMV-secreting high-quality monoclonal antibody hybridoma cell lines and the establishment of serological detection methods for the virus provide material and technical support for the diagnosis and detection, epidemiological investigation, and scientific prevention and control of ToMMV.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell strain secreting tomato mottle mosaic virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Tomato mottle mosaic virus (ToMMV) is a new species of Tobamovirus in the family Viraviridae, and its genome is positive-sense single-stranded RNA. The genome of ToMMV contains four open reading frames. Like tobacco mosaic virus, it expresses four proteins, encoding a 126kD and a 183kD replication-required protein, and also encoding a 30kD motor protein and a 17.5kD coat protein. In 2013, the new virus was first discovered on Mexican tomatoes, and then the virus was detected on tomatoes in the United States, Israel, Brazil, Spain and other places. Tomatoes in Yunnan and Hainan provinces in my country were also infected by the virus. [0003] ToMMV can harm a variety of Solanaceae crops, and the main symptoms of the disease ar...

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Problems solved by technology

However, the establishment of serological methods depends on specific high-quality virus antibodies

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