Development of a blocking ELISA for detection of Mycoplasma            hyopneumoniae infection based on a monoclonal antibody against protein          P65

Maojun LIU; Gaimei DU; Yue ZHANG; Yuzi WU; Haiyan WANG; Bin LI; Yun BAI; Zhixin FENG; Qiyan XIONG; Fangfang BAI; Glenn F BROWNING; Guoqing SHAO

doi:10.1292/jvms.15-0438

Immunology

Development of a blocking ELISA for detection of Mycoplasma hyopneumoniae infection based on a monoclonal antibody against protein P65

Maojun LIU, Gaimei DU, Yue ZHANG, Yuzi WU, Haiyan WANG, Bin LI, Yun BAI, Zhixin FENG, Qiyan XIONG, Fangfang BAI, Glenn F BROWNING, Guoqing SHAO

Author information

Maojun LIU
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences. Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture. National Center for Engineering Research of Veterinary Bio-Products, Nanjing 210014, China
Key Laboratory of Food Quality and Safety of Jiangsu Province-State Key Laboratory Breeding Base, Nanjing 210014, China
Gaimei DU
Department of Animal Science and Technology, Jinling Technology Institution, Nanjing 210038, China
Yue ZHANG
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences. Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture. National Center for Engineering Research of Veterinary Bio-Products, Nanjing 210014, China
Yuzi WU
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences. Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture. National Center for Engineering Research of Veterinary Bio-Products, Nanjing 210014, China
Haiyan WANG
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences. Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture. National Center for Engineering Research of Veterinary Bio-Products, Nanjing 210014, China
Bin LI
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences. Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture. National Center for Engineering Research of Veterinary Bio-Products, Nanjing 210014, China
Yun BAI
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences. Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture. National Center for Engineering Research of Veterinary Bio-Products, Nanjing 210014, China
Zhixin FENG
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences. Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture. National Center for Engineering Research of Veterinary Bio-Products, Nanjing 210014, China
Qiyan XIONG
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences. Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture. National Center for Engineering Research of Veterinary Bio-Products, Nanjing 210014, China
Fangfang BAI
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences. Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture. National Center for Engineering Research of Veterinary Bio-Products, Nanjing 210014, China
Glenn F BROWNING
Asia-Pacific Centre for Animal Health, Faculty of Veterinary and Agricultural Sciences, University of Melbourne, Parkville, Victoria 3010, Australia
Guoqing SHAO
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences. Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture. National Center for Engineering Research of Veterinary Bio-Products, Nanjing 210014, China
Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225009, China

Keywords: blocking ELISA, Mycoplasma hyopneumoniae, P65

JOURNAL FREE ACCESS

2016 Volume 78 Issue 8 Pages 1319-1322

DOI https://doi.org/10.1292/jvms.15-0438

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Abstract

Mycoplasma hyopneumoniae causes porcine enzootic pneumonia, an economically important disease of swine. A more sensitive and reliable method for detection of serum antibodies is needed for epidemiological investigations and to evaluate the effect of immunization. We expressed the M. hyopneumoniae protein P65 in Escherichia coli and produced a monoclonal antibody (mAb) that bound specifically to recombinant P65. Using this mAb, a blocking enzyme linked immunosorbent assay (ELISA) was developed. The blocking ELISA had similar specificity to and sensitivity with the commercial ELISA produced by IDEXX. Thus, this blocking ELISA is a useful test for serological confirmation of M. hyopneumoniae infection.

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