[Establishment of sandwich ELISA for detecting Staphylococcal enterotoxin B in different matrices].
ABSTRACT To establish sandwich ELISA for the detection of Staphylococcal enterotoxin B(SEB) and characterize the sensitivity and specificity of it in different materials.
The anti-SEB monoclonal antibodies (mAb) B4 and D6 were employed as capture antibody and detecting antibody respectively after being purified by Q sepharose fast flow chromatography and horseradish peroxidase(HRP) conjugation.
The sensitivity of this assay reached 0.2 ng of SEB per mL of PBS with BSA and fetal bovine serum. It reached 0.39 ng of SEB per mL of 50 g/L skim milk, human urine and water. The concentration curve generated from SEB standard curve was linear with the range of 0.78-12.5 microg/L, (r(2)=0.99). This assay was highly reproducible and the coefficient of variation(CV) was less than 10%.No cross-reactivity between SEA and SECl was found.
This assay offers a viable method with high sensitivity and specificity for detecting SEB and it may be used for SEB detection in clinical application and food samples.