Abstract: 8-week-old female BALB/c mice were immunized with Mycoplasma bovis HB0801. By using the hybridoma technique, six hybridoma cell lines secreting monoclonal antibodies (MAbs) were identified. The ascite for the six MAbs were prepared and the characteristics of them were further studied. All of the six MAbs belonged to IgG category. The antibody titers of these MAbs ranged from 1?10⁵ to 1.6?10⁶. The specificity of these MAbs was detected by ELISA. The result suggested that all these MAbs can react with Mycoplasma bovis strains clinically isolated from China and the ATCC reference strain PG45, but they did not react with other bovine pathogenic bacteria tested such as Pasteurella multocida, Arcanobacterium pyogenes and so on. All MAbs investigated cross-reacted with Mycoplasma agalactiae which was known to be closely related to Mycoplasma bovis, prevalent in goats but not in cattle. Among them, two MAbs 1A5 and 1C11 did not exhibit further cross-reactions to other Mycoplasma species tested. In addition, Western blotting assay demonstrated that these six antibodies recognized various Mycoplasma bovis proteins. In conclusion, the MAbs prepared in this study layed the foundation for the establishment of novel detection methods for Mycoplasma bovis and studying its pathogenesis.

Key words: Mycoplasma bovis; monoclonal antibodies (MAbs); Western blotting; ELISA; hybridoma; specificity