Preparation and identification of a monoclonal antibody against Mycoplasma hyorhinis / 中国人兽共患病学报

ABSTRACT

The aim of this study was to produce a monoclonal antibody against Mycoplasma hyorhinis,which would be useful in the diagnosis and pathogenesis researches of M.hyorhinis.BALB/c mice were immunized with whole cell protein of M.hyorhinis,and then the monoclonal antibody (McAb) was prepared by hybridoma technique.The isotype of the McAb was identified,and then the titer and specificity were analyzed.The prepared McAb was used to detect M.hyorhinis in the colony immunoboltting assay and indirect immunofluorescence assay.A hybridoma cell line was obtained,and was named Mhr-08.The McAb belonged to IgG1 isotype,and the light chain was κ type.The titer was detected to be 1 ∶ 102 400 by ELISA.The result of Western-blot indicated that this McAb strongly reacted to a 43 kDa protein of M.hyorhinis,without cross-reaction with other swine mycoplasmas,Escherichia coli or KM2 medium.It revealed that the McAb recognized a surface membrane protein and was successfully applied to detect M.hyorhinis in colony immunoboltting assay.The mycoplasmas bound to the tracheal epithelial cells was successfully detected by using this McAb in indirect immunofluorescence assay.In conclusion,a McAb against M.hyorhinis was successfully prepared,which provides as a useful tool for the diagnosis and pathogenesis researches.