Production of monoclonal antibodies specific to avian pneumoviruses

Monoclonal antibodies (MAbs) specific to avian pneumoviruses were obtained for the diagnosis of turkey rhinotracheitis (TRT) and swollen head syndrome (SHS). The vaccinal BUT strain of TRT virus was used as an antigen for mouse immunisation. The virus was propagated in a Vero cell culture followed by purification and concentration by ultracentrifugation. The obtained antigen was used for threefold Balb/c mouse immunisation. The spleen from a mouse with the highest anti-TRT antibody level was sampled. Fusion of splenic cells with Sp2/0-Ag 14 myeloma cells was conducted in the presence of PEG-1450. The ELISA and peroxidase-linked assay (PLA) were used to detect antibodies specific to TRT virus in the sera of immunised mice, in the supernatant of hybridoma cell cultures and in the ascitic fluid of mice. The antigenic specificity of MAbs was determined by the immunoblotting technique and their isotype by agar gel immunodifusion method and ELISA. Seven secretive clones were obtained from 2 fusions. The MAbs produced by the clones were detected by the PLA and in the case of 3 clones the ELISA was used. One clone secreted the IgG2a and the remaining clones the lgG1 immunoglobulines. All MAbs recognised the surface G glycoprotein. The MAbs obtained were successfully used to distinguish the Polish avian pneumovirus APV isolates and to detect the virus in the tissues of the respiratory system of chickens infected experimentally.