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目的制备一种表面包被有大肠埃希菌O157:H7多克隆抗体的纳米级葡聚糖免疫磁颗粒,在检测大肠埃希菌O157:H7的同时,对磁颗粒的应用条件进行优化。方法FeCl3和FeCl2在氨水条件下与葡聚糖反应生成纳米葡聚糖磁颗粒,利用高碘酸钠将其氧化后,在表面包被上大肠埃希菌O157:H7多克隆抗体,制成纳米免疫磁颗粒进行检测。结果该方法可在15 min内完成对样品的分离,检测限为101CFU/ml甚至更少,当样品中含有108CFU/ml的杂菌时,检测限为101~102CFU/ml,杂菌对检测结果影响甚微。结论利用纳米级葡聚糖免疫磁颗粒分离目的菌是一种灵敏度高、特异性强、操作简便的有效方法。
OBJECTIVE To prepare a nano-sized dextran immunomagnetic particle coated with the Escherichia coli O157: H7 polyclonal antibody on the surface of Escherichia coli O157: H7, and optimize the conditions of application of the magnetic particle. Methods FeCl3 and FeCl2 reacted with dextran in aqueous ammonia to produce nano-dextran magnetic particles, which were oxidized with sodium periodate and then coated with the Escherichia coli O157: H7 polyclonal antibody on the surface to form nanometer Immunomagnetic particles for testing. Results The method can complete the separation of samples within 15 min with a detection limit of 101 CFU / ml or less. When the sample contains 108 CFU / ml of bacteria, the detection limit is 101-102 CFU / ml. Little effect. Conclusion The use of nano-dextran immunomagnetic particles isolated from the target bacteria is a high sensitivity, specificity and easy to operate an effective method.