Establishment and application of a rapid detection method for Escherichia coli O157∶H7 in beef
ZHAO Ya-nan1, ZENG De-xin1, GUO De-hua2, JIANG Yuan2, JIANG Lu-yan3, LI Xin-ni4, CHEN Wei4, TANG Fang1, XUE Feng1, DAI Jian-jun1
1. International Cooperative Laboratory for Animal Health and Food Safety, Nanjing Agricultural University, Nanjing 210095, China; 2. Shanghai Customs Animal and Plant and Food Inspection and Quarantine Technology Center, Shanghai 200135, China; 3. Nanjing Customs Animal and Plant and Food Testing Center, Nanjing 210001, China; 4. College of Food and Bioengineering, Hefei University of Technology, Hefei 230601, China
Abstract：To establish a rapid and accurate method for the detection of Escherichia coli O157∶H7 in beef. The anti-E. Coli O157∶H7 monoclonal antibody 2G5 as capture antibody and enzyme-labeled monoclonal antibody 2E3 (HRP-2E3) as detection antibody were used to establish a double-antibody sandwich ELISA method for detecting E. coli O157∶H7. Then, the reaction conditions and the method were optimized. The optimal coating concentration of monoclonal antibody 2G5 was 4.48 μg/mL, and the optimal detection concentration of HRP-2E3 was 19.9 μg/mL . The limit of detection (LOD) of E. coli O157∶H7 reached 105 CFU/mL, showing in good sensitivity. Also, this method had no cross-reaction with other Escherichia coli, Salmonella, Listeria, etc, which showed high specificity. Variable coefficient of intra-and inter-plate were all less than 7% showing high precision. LOD of E. coli O157∶H7 in artificial contamination of beef samples was 1 CFU/25g after enrichment for 8h. Thus, the established sandwich ELISA method has good sensitivity, specificity and reproducibility. It can be used for the detection of clinical actual samples.
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2020, Vol. 36 
