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Objective To prepare a high-titer monoclonal antibody detection kit for aflatoxin B1 (AFB1) and rapidly and accurately detect AFB1 in food. Methods By carbodiimide method, AFB1 was conjugated with bovine albumin (BSA) to form immune antigen, and with chicken ovalbumin (OVA) to prepare detection antigen. AFB1 monoclonal antibody was prepared by immunizing mice, cell fusion, screening of hybridoma cells, inducing ascites in vivo, isolation and purification, and a kit for rapid detection of AFB1 was developed. Results The prepared monoclonal antibody had a titer of up to 1:27 w and showed weak cross-reactivity to Shentuqumycin, with a reaction rate of 13%. No cross-reactivity was observed with the other 2 compounds. The relative standard deviation of the intra- and inter-batch tests for the spiked samples was 2.6%-3.6% and 5.0%-8.0%, respectively. The recoveries were 89.82%-103.64%. The detection sensitivity (median inhibitory concentration value) was 650 pg/mL, with a detection range of 156-5000 pg/mL with the limit of detection of 100 pg/mL. When AFB1 content was detected in corn flour from different origins, the detection results were 9.756, 2.483, 3.995, 39.080 and 7.831 μg/kg. Conclusion This research has prepared high-titer monoclonal antibodies against AFB1. These antibodies can not only be utilized for the development of test kit detection experimental methods but also lay the foundation for the development of rapid test strips based on AFB1 monoclonal antibodies and the colloidal gold method.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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