免疫磁珠结合显色平板快速检测婴儿配方奶粉中阪崎克罗诺杆菌  

Rapid Detection of Cronobacter sakazakii in Powdered Infant Formula by Immunomagnetic Beads Combined with Chromogenic Plate

作　　者：王磊 张洁 张文馨 崔晓雅 WANG Lei;ZHANG Jie;ZHANG Wenxin;CUI Xiaoya(Tianjin Institute of Food Safety Inspection Technology,Tianjin 300308,China)

机构地区：[1]天津市食品安全检测技术研究院,天津300308

出　　处：《食品研究与开发》2023年第14期187-191,204,共6页Food Research and Development

摘　　要：该研究利用羧基化磁珠偶联兔多克隆抗体,制备特异性免疫磁珠,并优化制备条件,然后与显色培养基联用,建立免疫磁珠-显色平板检测方法,用于富集分离婴儿奶粉中的阪崎克罗诺杆菌。结果表明,磁珠与抗体的最佳偶联温度37℃、偶联时间2.0 h,1 mg磁珠的最佳抗体添加量为200μg,饱和偶联量为182.77μg。制得的免疫磁珠,捕获目标菌的浓度范围为10~2~10~7 CFU/mL,捕获率为68.4%~82.4%,与非目标菌反应率低于10%。检测方法应用于模拟污染样品,在目标菌浓度10~2~10~5 CFU/mL时,捕获率为66.2%~71.5%。相比传统方法,该方法的检测周期缩短了2~3 d。This study used carboxylated magnetic beads coupled with rabbit polyclonal antibodies to prepare specific immunomagnetic beads.The preparation conditions were optimized,and they were then used in conjunction with a chromogenic medium to establish an immunomagnetic beads-chromogenic plate detection method for the enrichment and separation of Cronobacter sakazakii in powdered infant formula.The results showed that the optimal coupling temperature between the magnetic beads and antibodies was 37 ℃,with a coupling time of 2.0 h.The optimal antibody dosage for 1 mg of magnetic beads was 200 μg,and the saturation coupling amount was 182.77 μg.The prepared immunomagnetic beads had a concentration range for capturing the target bacteria of 10~2 to 10~7 CFU/mL,with a capture efficiency of 68.4%-82.4%,and a reaction rate with non-target bacteria of less than 10%.The detection method was applied to simulated contaminated samples,and at target bacterial concentrations ranging from 10~2 to 10~5 CFU/mL,the capture efficiency was 66.2%-71.5%.Compared with traditional methods,this method reduced the detection time by 2-3 d.

关 键 词：阪崎克罗诺杆菌 免疫磁珠 显色培养基 快速检测 婴儿配方食品 

分 类 号：TS252.51[轻工技术与工程—农产品加工及贮藏工程]