Preparation of colloidal gold-labeled strip of monoclonal antibody against BP subunit of PI-2a pili islands of bovine mastitis Streptococcus agalactiae
BU Ri-e1, 2, 3, WU Jin-hua1, 2, 3, XI Lin-gao-wa1, 2, 3, CHEN Jin-long4, WANG Jin-liang4
1.Inner Mongolia Autonomous Region Engineering Technology Research Center of Prevention And Control the Pathogenic Bacteriain Milk,Tongliao 028043,China; 2.College of Life Science, Inner Mongolia University for Nationalities,Tongliao 028043,China; 3.Research Institute for Pathogenic in Milk of Inner Mongolia University for Nationalities,Tongliao 028043,China; 4.Shandong Lvdu Bio-sciences and Technology Limited company, Binzhou 256600,China
Abstract：In this experiment, by optimizing performance of the monoclonal antibody against PI-2a pili island BP subunit of Streptococcus agalactiae, the colloidal gold labeling for the detection of S.agalactiae of mastitis was prepared. The mice were firstly immunized with purified protein expressed from pET30a (+)-BP recombinant plasmid. Then mice spleen cells were taken aseptically for fusion with sp2/0 myeloma cells. The hybridoma cells were screened and monoclonal antibodies were prepared. The monoclonal antibody was labeled with colloidal gold for the detection kit preparation, and its sensitivity and specificity were evaluated. After screening, a positive hybridoma cell line was obtained, and the antibody titer reached 1∶256 00. The sensitivity measurement of colloidal gold marking strip reached 106 CFU / mL. Specificity test results showed that there was no positive reaction between colloidal gold label test strip and bovine milk samples artificially contaminated by S. aureus, Escherichia coli, Salmonella paraturium, Mycobacterium paratuberculosis and Streptococcus suppurate. The positive results were only found in the milk samples of Streptococcus agalactiea. In conclusion, the colloidal gold labeled immune-chromatographic strip has practical application value and will provide a new and convenient technical product for the screening and detection of bovine mastitis.
布日额, 吴金花, 锡林高娃, 陈金龙, 王金良. 牛乳腺炎无乳链球菌PI-2a菌毛岛屿BP亚基单克隆抗体胶体金标记试纸条制备[J]. 中国人兽共患病学报, 2020, 36(1): 45-49.
BU Ri-e, WU Jin-hua, XI Lin-gao-wa, CHEN Jin-long, WANG Jin-liang. Preparation of colloidal gold-labeled strip of monoclonal antibody against BP subunit of PI-2a pili islands of bovine mastitis Streptococcus agalactiae. Chinese Journal of Zoonoses, 2020, 36(1): 45-49.
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