荧光微球免疫层析技术定量检测河鲀毒素

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。

Microsphere-Based Fluorescence Immunochromatographic Assay for Quantitative Detection of Tetrodotoxin

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 from 2 to 200 ng/mL for the detection of tetrodotoxin (y = ?0.15lnx + 0.95, R2 = 0.98) with an IC50 of 18.4 ng/mL. The limit of detection (LOD) for tetrodotoxin was 10 μg/kg in puffer with an average relative standard deviation (RSD) of less than 25% (n = 6). The assay method could be applied on heat-processed products without the need for large-scale equipment. The whole analysis process took only 15 min for each sample.