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Background   Mycoplasma bovis  ( M. bovis ) is a major, but often overlooked, pathogen documented to cause respiratory disease, mastitis, and arthritis in cattle throughout China since 2008. Here, we report the development of a direct competitive enzyme-linked immunosorbent assay (Dc-ELISA) to detect  M. bovis  antibody.    Results  We used a recombinant P48 protein and monoclonal antibody (mAb) 10E. MAb 10E, prepared against the recombinant P48 protein of  M. bovis , identified all  M. bovis  strains with no cross-reactivity with other related pathogens. Coating micro plates with P48 protein instead of whole  M. bovis  cells as well as the use of mAb 10E produced a specific and sensitive Dc-ELISA for  M. bovis  antibody detection with a cut-off percent inhibition (PI) value of 32%. Compared with two commercial indirect ELISA (i-ELISA) kits, our Dc-ELISA offered a higher positive detection rate in 165 clinical bovine serum samples.    Conclusions  A rapid, sensitive, and reliable serological diagnosis method was developed for  M. bovis , which can facilitate  M. bovis  surveillance, assisting researchers in understanding the ecology and epidemiology of  M. bovis .

Development of a direct competitive ELISA for the detection of Mycoplasma bovisinfection based on a monoclonal antibody of P48 protein