免疫磁珠法富集沙门氏菌的优化及应用  

Optimization and application of immunomagnetic beads for enriching Salmonella spp

作　　者：山珊[1] 牛瑞江[1] 赖卫华[1] 刘道峰[1] 魏华[1] 熊勇华[1] 

机构地区：[1]南昌大学食品科学与技术国家重点实验室,南昌大学,江西南昌330047

出　　处：《食品工业科技》2013年第13期153-156,共4页Science and Technology of Food Industry

基　　金：国家科技支撑计划(2011BAK10B01-03);江西科技厅社会发展攻关项目(20111BBG70010-3)

摘　　要：采用活泼酯法将磁珠和抗体偶联,制备沙门氏菌免疫磁珠,系统地优化了偶联条件和免疫磁珠捕获工艺。结果表明,最佳偶联条件:每毫克磁珠抗体偶联量为50μg,免疫磁珠的添加量为0.4mg,菌液浓度为102～106CFU/mL时,捕获效率为65%～82%;免疫磁珠捕获工艺:目的菌最佳孵育时间为45min,最佳磁分离时间为3min,磁场强度为0.8T,磁珠粒径为180nm;对鼠伤寒沙门氏菌、猪霍乱沙门氏菌、甲型副伤寒沙门氏菌、鸭沙门氏菌和肠炎沙门氏菌的捕获效率分别为61.4%、45.8%、48.2%、78.5%、64.1%,对鸡蛋清、蛋黄、牛奶和纯培养基中沙门氏菌捕获效率分别为44.3%、43.2%、65.2%和72.5%。免疫磁分离富集技术具有分离速度快、操作简单、捕获效率高、可重复性好等特点,可在食品样品预处理中广泛应用。Anti-Salmonella polyclonal antibody was coupled to magnetic beads using EDC-NHSS method,and the coupling conditions and immunomagnetic beads capture process were optimized in this paper.The result showed that the optimum conditions were as follows.-501Jg antibody in ling magnetic beads; the amount of immunemagnetic beads was 0.4mg.The capture efficiency of the immunomagnetic beads for Salmonella spp.were from 65% to 82% when the concentration in the range of 102-106CFU/mL.Incubation time and separation time was 45rain and 3min,respectively.The magnetic density was 0.8T and the diameter of beads was 180rim.The capture efficiency with Salmonella typhimurium, Salmonella choleraesuis, Salmonella parayohi A, Salmonella anatum and Salmonella enteritidis were 61.4%, 45.8%, 48.2%, ？8.5% and 64.1%, respectively. The capture efficiency of the immunomagnetic beads for Salmonella spp. in egg white, yolk, milk, and medium were 44.3%, 43.2%, 65.2%, and ？2.5%,respectively. Immunomagnetic separation and enrichment technique has high separration speed, simple operation,high capture efficiency,good repeatability,so it would be widely used for food sample pretreatment.

关 键 词：免疫磁分离 沙门氏菌 捕获效率 

分 类 号：TS201.3[轻工技术与工程—食品科学]