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Monoclonal Antibody-Based Sandwich ELISA for the Detection of Staphylococcal Enterotoxin A
State Key Laboratory of Food Science & Technology, School of Food Science & Technology, Jiangnan University, Wuxi 214122, China
Research Centre of Hunan Entry-Exit Inspection and Quarantine Bureau, Changsha 410001, China
These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 19 March 2013; in revised form: 26 March 2013 / Accepted: 15 April 2013 / Published: 19 April 2013
Abstract: A sensitive and specific monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (ELISA) was established and validated for the detection of staphylococcal enterotoxin A (SEA). After routine fusion and selection, 10 monoclonal antibodies showed high affinity for SEA. An optimal pair for sandwich ELISA was selected by pairwise interaction analysis. After optimization, the limit of detection (LOD) and linear dynamic range of the method were established, and were found to be 0.0282 ng/mL and 0.06–2 ng/mL, respectively. The recovery in pure milk ranged from 82.67% to 111.95% and the intra- and inter-assay coefficients of variation ranged from 3.16% to 6.05% and from 5.16% to 10.79%, respectively. Cross-reactivity with staphylococcal enterotoxin B (SEB), staphylococcal enterotoxin C (SEC), staphylococcal enterotoxin D (SED), and staphylococcal enterotoxin E (SEE) in this method were insignificant. These results indicate that the sandwich ELISA method developed in our study is effective for routine identification of SEA in food samples.
Keywords: staphylococcal enterotoxin A (SEA); monoclonal antibody; sandwich ELISA; detection
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MDPI and ACS Style
Kuang, H.; Wang, W.; Xu, L.; Ma, W.; Liu, L.; Wang, L.; Xu, C. Monoclonal Antibody-Based Sandwich ELISA for the Detection of Staphylococcal Enterotoxin A. Int. J. Environ. Res. Public Health 2013, 10, 1598-1608.
Kuang H, Wang W, Xu L, Ma W, Liu L, Wang L, Xu C. Monoclonal Antibody-Based Sandwich ELISA for the Detection of Staphylococcal Enterotoxin A. International Journal of Environmental Research and Public Health. 2013; 10(4):1598-1608.
Kuang, Hua; Wang, Wenbing; Xu, Liguang; Ma, Wei; Liu, Liqiang; Wang, Libing; Xu, Chuanlai. 2013. "Monoclonal Antibody-Based Sandwich ELISA for the Detection of Staphylococcal Enterotoxin A." Int. J. Environ. Res. Public Health 10, no. 4: 1598-1608.