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Open Access Methodology article

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

Ping Fu1, Zhenhong Sun1, Yuewei Zhang1, Ziqiang Yu1, Haiyan Zhang1, Dan Su2, Fei Jiang1 and Wenxue Wu1*

Author Affiliations

1 Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing, China

2 Shandong Vocational Animal Science and Veterinary College, Weifang, China

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BMC Veterinary Research 2014, 10:42  doi:10.1186/1746-6148-10-42

Published: 18 February 2014

Abstract

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.

Keywords:
Direct competitive ELISA; Mycoplasma bovis; Monoclonal antibody