摘要
建立河鲀毒素的荧光微球免疫层析检测方法。基于抗体亲和位点保护标记方法制备抗河鲀毒素抗体偶联荧光微球,方法简单稳定,所有结合分离步骤均在亲和柱中完成,避免了亲和位点被破坏而且荧光信号更强,将检测灵敏度提高至原来的8倍左右。所建立的河鲀毒素检测方法IC50为18.4 ng/m L,线性范围为2~200 ng/m L(y=-0.15ln x+0.95,R2=0.98)。检测河鲀样本的最低检出限为10μg/kg,平均相对标准偏差小于25%(n=6)。该方法适用于热加工样品的检测,整个检测过程仅需15 min,且无需使用大型设备,为河鲀餐前速测提供了一定技术手段。
A microsphere-based fluorescence immunochromatographic assay was established for the detection of tetrodotoxin.A binding site protection procedure was developed for antibody labeling with fluorescent microspheres,which prevented damage to the antibody binding site and consequently enhanced the fluorescence signal.The whole process of binding and separation was efficiently completed on an affinity spin column.The sensitivity of the antibody-microsphere conjugate was nine times higher than that prepared by direct labeling.The immunochromatographic assay exhibited a linear range from2to200ng/mL for the detection of tetrodotoxin(y=?0.15lnx+0.95,R2=0.98)with an IC50of18.4ng/mL.The limit of detection(LOD)for tetrodotoxin was10μg/kg in puffer with an average relative standard deviation(RSD)of less than25%(n=6).The assay method could be applied on heat-processed products without the need for large-scale equipment.The whole analysis process took only15min for each sample.
作者
张世伟
王士峰
姚添琪
杨国武
赖心田
ZHANG Shiwei;WANG Shifeng;YAO Tianqi;YANG Guowu;LAI Xintian(Shenzhen Academy of Metrology and Quality Inspection, Shenzhen 518102, China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2017年第20期312-317,共6页
Food Science
基金
深圳市科技计划项目(JCY20150626104807916)
关键词
河鲀毒素
亲和位点保护
荧光微球
免疫层析
tetrodotoxin
binding site protection
fluorescent microspheres
immunochromatography
