大田软海绵酸和鳍藻毒素时间分辨荧光免疫层析试纸条的研制与应用

Development and application of time-resolved fluorescent immunochromatography test strip for okadaic acid and dinophysis toxins

腹泻性贝类毒素主要包括大田软海绵酸(okadaic acid,OA)、鳍藻毒素-1(dinophysis toxin-1,DTX-1)和鳍藻毒素-2(dinophysis toxin-2,DTX-2)。被贝类毒素污染的海产品可对食用者的健康构成危害。在我国近岸海域OA常有检出,DTX-1和DTX-2偶有检出。本文用荧光免疫微球标记抗OA单克隆抗体,利用牛血清白蛋白合成高偶联比的包被抗原,以硝酸纤维素膜为载体,运用免疫层析技术,研制了能够同时快速准确检测OA、DTX-1和DTX-2的时间分辨荧光免疫层析试纸条,定量限分别为90.6μg/kg、115.4μg/kg和69.8μg/kg。实验验证结果表明:本文研制的时间分辨荧光免疫层析试纸条对含有OA、DTX-1和DTX-2中的一种、两种或三种组分的毒素样品检测结果,与液相色谱-串联质谱法检测结果的相关性良好。该时间分辨荧光免疫层析试纸条灵敏、准确、操作简便,成本低,在我国海洋环境监测及海产品食用安全检测中,应用前景良好。

Diarrhetic shellfish poisoning(DSP)are mainly composed of okadaic acid(OA),dinophysis toxin-1(DTX-1)and dinophysis toxin-2(DTX-2),which are seriously harmful for human consuming seafood.OA has been often found,while DTX-1 and DTX-2 been occasionally detected at the coastal waters of China.This paper mainly introduced the principle of immunochromatographic assay,and used nitrocellulose membrane as carrier.Fluorescent immunobeads were adopted to label monoclonal antibodies against okadaic acid.The timeresolved fluorescent immunochromatography strip for rapid and accurate detection of OA,DTX-1 and DTX-2 was developed by using bovine serum albumin(BSA)to synthesize high coupling ratio coated antigen.The time-resolved fluorescent immunochromatography strip limits of quantitation for OA,DTX-1 and DTX-2 were 90.6μg/kg,115.4μg/kg and 69.8μg/kg,respectively.The experimental results show that the time-resolved fluorescent immunochromatography strip developed in this paper has a good correlation with the results of LC-MS/MS for the detection of toxin samples containing one,two or three components of OA,DTX-1 and DTX-2.The time-resolved fluorescent immunochromatographic strip is sensitive,accurate,easy to operate and low cost.It has a good application prospect in marine environmental monitoring and seafood food safety detection in China.