The invention discloses a hybridoma cell strain secreting a monoclonal antibody against banana bunchy top virus, and an application of the monoclonal antibody thereof. A BALB / c mouse is immunized with purified banana bunchy top virus (BBTV) virions as antigens, and a hybridoma cell strain 22E3 which can be subcultured stably and secretes a monoclonal antibody against BBTV is obtained through cell fusion, screening, and cloning, and the accession number is CGMCC No. 8780. The 22E3 monoclonal antibody has an ascetic indirect ELISA titer of up to 10<-7>, and the antibody type and subtype are IgG1, kappa chain. Specific reaction can be carried out between the monoclonal antibody and BBTV. A dot-ELISA method for detecting BBTV in bananas established with the 22F3 monoclonal antibody can detect the virus even when a diseased leaf is diluted by 1:320 times (w / v, g / mL). The acquisition of the hybridoma cell secreting monoclonal antibody against BBTV and its monoclonal antibody, and the establishment of a related serological detection method provide technical and material support for the diagnosis, prediction and scientific prevention and control of the viral disease.