Hybridoma cell strain secreting monoclonal antibody resistant to Candidatus Liberibacter asiaticus pathogens and application of monoclonal antibody

Abstract

The invention discloses a hybridoma cell strain secreting a monoclonal antibody resistant to Candidatus Liberibacter asiaticus pathogens and an application of the monoclonal antibody. A Candidatus Liberibacter asiaticus vein crude extract is taken as an antigen to immunize a BALB / c mouse, one hybridoma cell strain 11H9 capable of secreting the monoclonal antibody resistant to Candidatus Liberibacter asiaticus pathogens is obtained by cell fusion, screening and cloning, and the preservation number of the hybridoma cell strain is CGMCC No.17285. Indirect ELISA valence of monoclonal antibody ascites secreted by the cell strain reaches 10<-7>, the type and the subclass of the antibody are IgG1 and kappa light chains, and the monoclonal antibody has a specific reaction with Candidatus Liberibacter asiaticus pathogen protein of 56 kDa in sick leaves and avoids immunoreaction with healthy leaves. ACP-ELISA, dot-ELISA and Tissue blot-ELISA detection methods for detecting the Candidatus Liberibacter asiaticus pathogens are established on the basis of the monoclonal antibody11H9, wherein sensitivity of the ACP-ELISA and dot-ELISA methods for detecting the Candidatus Liberibacter asiaticus vein crude extract reaches 20480-fold dilution and 10240-fold dilution (w / v,g / mL). The material and technical support is provided for diagnosis and detection of Candidatus Liberibacter asiaticus, epidemiological analysis, sterile seedling production and scientific prevention and control through establishment of the preparation and detection methods of the monoclonal antibody resistant to the Candidatus Liberibacter asiaticus pathogens.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell line secreting monoclonal antibody against citrus huanglongbing bacteria and the application of the monoclonal antibody. Background technique [0002] At the beginning of the 20th century, citrus huanglongbing broke out in more than 50 countries and regions in Asia, Africa, Oceania, South America and North America. Citrus huanglongbing was reported for the first time in South my country in 1919 in China. At present, the disease has been prevalent in Guangdong, Jiangxi, Hainan, Guangxi, Fujian, Zhejiang, Yunnan, Guizhou, Hunan, Chongqing and other provinces (autonomous regions or municipalities directly under the central government). The citrus cultivation areas are all affected by citrus greening disease, the affected area has reached 80% of the total area, and the output has lost 85%. The citrus industry is facing an unprecedented crisis due to the widespread outbre...

AI Technical Summary

Problems solved by technology

[0003] At present, low-efficiency symptom observation, indicator plants, electron microscope observation, PCR and other methods are mostly used in the detection and diagnosis of citrus Huanglongbing bacteria. There is no fast, practical and high-throughput detection technology and method

Images