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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Affiliations:1: Huaian Entry-Exit Inspection and Quarantine Bureau, Huaian, 223001, China 2: School of Food Science & Technology, State Key Lab of Food Science & Technology, Jiangnan University, Wuxi, 214122, China