TY - JOUR ID - 3195 TI - Development of an indirect ELISA for bovine mastitis using Sip protein of Streptococcus agalactiae JO - Iranian Journal of Veterinary Research JA - IJVR LA - en SN - 1728-1997 AU - Bu, R. E. AU - Wang, J. L. AU - DebRoy, C. AU - Wu, J. H. AU - Xi, L. G. W. AU - Liu, Y. AU - Shen, Z. Q. AD - College of Life Science, Inner Mongolia University for Nationalities, Tongliao 028043, China AD - Shandong Binzhou Animal Science and Veterinary Medicine Academy, Binzhou 256600, Shandong Province, China; E. coli Reference Center, Department of Veterinary and Biomedical Sciences, Pennsylvania State University, University Park 16803, Pennsylvania, USA AD - E. coli Reference Center, Department of Veterinary and Biomedical Sciences, Pennsylvania State University, University Park 16803, Pennsylvania, USA AD - Shandong Binzhou Animal Science and Veterinary Medicine Academy, Binzhou 256600, Shandong Province, China Y1 - 2015 PY - 2015 VL - 16 IS - 3 SP - 283 EP - 287 KW - Bovine mastitis KW - ELISA KW - Streptococcus agalactiae DO - 10.22099/ijvr.2015.3195 N2 - The sip gene encoding for a conserved highly immunogenic surface protein of Streptococcus agalactiae was amplified using polymerase chain reaction (PCR) and subcloned into prokaryotic expression vector pET32a (+) and expressed as a recombinant protein in E. coli BL21 (DE3). An indirect enzyme linked immunosorbent assay (ELISA) was developed using the purified Sip protein as a coating antigen, which could identify S. agalactiae specific antibody in sera. The coating antigen at a concentration of 3.125 μg/ml, serum diluted to 1:160, and HRP-conjugated secondary antibody concentration at 1:4000 was found to be most effectivein exhibiting positive result. The ELISA was found to be highly specific for S. agalactiae that may be used for the detection of the pathogen in mastitis cases, for epidemiological studies and for surveillance. UR - https://ijvr.shirazu.ac.ir/article_3195.html L1 - https://ijvr.shirazu.ac.ir/article_3195_d09a4712b0f745324f86f206c917bc6f.pdf ER -