Authors: Pan Xue Yongming Li Zhujun Zhang Aihua Fu Fei Liu Xiaoming Zhang Yuanjie Sun Lili Chen Boquan Jin Kun Yang
Publish Date: 2011/05/11
Volume: 174, Issue: 1-2, Pages: 167-174
Abstract
An enzymelinked immunosorbent assay a horseradish peroxidasecatalyzed fluorogenic reaction and chemiluminescence CL analysis have been combined to develop a sandwich ELISA for Staphylococcal enterotoxin B SEB using monoclonal antibodies for different epitopes of SEB The enzyme catalyzed reaction of 34hydroxyphenyl propionate with the urea complex of hydrogen peroxide produced a fluorescent dimer which was detected by chemiluminescence analysis The CL response to SEB is linear in the range from 60 to 564 pg mL−1 r = 09993 and the detection limit is 33 pg mL−1 S/N = 3 Intra and interassay coefficients of variation are 70 at three concentrations 24 96 and 384 pg mL−1 The method was applied to the analysis of SEB in serum lake water and milk samples The results compared well with those obtained by conventional ELISAsProcedures of the proposed method A sandwich ELISA for Staphylococcal enterotoxin B SEB using a pair of monoclonal antibodies that recognizes different epitopes of SEB After the ELISA procedure PHPPA is reacted with Hydrogen peroxideurea with catalysis by HRPconjugated antiSEB to produce PHPPA fluorescent Dimer which is detected by TCPO chemiluminescenceThis work was supported financially by the Fundamental Research Funds for the Central Universities Program No GK20091004 the National High–tech RD Program 863 Program of China No 2006AA02A237 and the National Natural Science Foundation of China No 30872371
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