Development of a monoclonal antibody-based ELISA to detect Escherichia coli O157:H7 |
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| Authors: | Min Feng Qianqian Yong Wenbing Wang Libing Wang Chuanlai Xu |
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| Institution: | 1. Huaian Entry-Exit Inspection and Quarantine Bureau, Huaian 223001, China;2. School of Food Science &3. Technology, State Key Lab of Food Science &4. Technology, Jiangnan University, Wuxi 214122, China;5. School of Food Science & |
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| Abstract: | A pair of monoclonal antibodies (mAb) from 10 murine hybridomas secreting Escherichia coli O157:H7 (E. coli O157:H7)-specific mAbs were selected for the development of the sandwich ELISA to detect E. coli O157:H7. On the basis of pairwise interaction analysis, mAb-1 was selected as a capture antibody while mAb-6 was used as a detection antibody. The buffer system which provided the greatest difference between the specific E. coli O157:H7-positive antigen and the negative control was chosen. This sandwich ELISA showed good linearity when the concentration of E. coli O157:H7 was in the range of 105–108 cfu/mL, and the sensitivity was 1×104 cfu/mL. With 8-h enrichment of bacteria, this ELISA was found to detect 0.4 cfu/g E. coli O157:H7 in artificially contaminated green tea samples. |
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| Keywords: | Escherichia coli O157:H7 sandwich ELISA green tea monoclonal antibody |
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