Establishment of indirect ELISA for bovine mastitis Streptococcus agalactiae based on rSip-PGK-FbsA fusion protein
Streptococcus agalactiae is the one of the important caused agent for mastitis in cow.To establish a rapid and sensitive assay for detection of the antibody against S.agalactiae,a indirect ELISA was developed with prokaryotic expressed three proteins (rSip-PGK-FbsA) of S.agalactiae as the coating antigen.The reacted condition was optimized,including 2 μg/mL of coating rSip-PGK-FbsA,the 1∶160 dilution of the serum to be detected,and detecting with 1∶6,000 dilution of IgG-HRP.The results of specificity,sensitivity and repeatability tests showed that the assay had no reactions with the positive serum of S.pyogenes,E.coli,S.aureus and S.epidermidis.The positive coincidence rate of this method was 98.6％,higher than that ELISA method coating with the SIP and PGK protein.This method had high sensitivity.The detection rate was 46.53％ of 389 clinical samples.This indirect ELISA method is a more accurate and reliable serological method for the rapid detection of antibody against S.agalactiae.
NETL
NSTL
万方数据
维普
