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[Establishment of sandwich ELISA for detecting Staphylococcal enterotoxin B in different matrices]
Qi Li1, Li-jie Chen, Bang-quan Dong
1Department of Immunology, Fourth Military Medical University, Xi'an 710032, China.
Aim:
To establish sandwich ELISA for the detection of Staphylococcal enterotoxin B(SEB) and characterize the sensitivity and specificity of it in different materials.
Methods:
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.
Results:
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.
Conclusion:
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.
