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用于检测吗啡的量子点标记抗体荧光免疫分析方法的开发

Development of Quantum Dots-Labeled Antibody Fluorescence Immunoassays for the Detection of Morphine.

作者信息

Zhang Can, Han Yufeng, Lin Li, Deng Nannan, Chen Bo, Liu Yuan

机构信息

School of Food & Biological Engineering, Jiangsu University , Zhenjiang 212013, P. R. China.

Zhenjiang Institute for Drug Control, Zhenjiang 212000, P. R. China.

出版信息

J Agric Food Chem. 2017 Feb 15;65(6):1290-1295. doi: 10.1021/acs.jafc.6b05305. Epub 2017 Feb 6.

Abstract

Quantum dots (QDs)-labeled antibody fluorescence immunoassays (FLISA) for the detection of morphine were developed. Quantum dots (CdSe/ZnS), which contained carboxyl, were used to label antimorphine antibody by 1-ethyl-3-(3-dimethylaminoprophyl) carbodiimide hydrochloride/N-hydroxysulfosuccinimide, which were used as coupling reagents. The CdSe/ZnS QDs labeled antimorphine antibody (QDs labeled Ab) was characterized by fluorescence spectrum and gel electrophoresis. Plate-based FLISA and nitrocellulose membrane-based flow-through FLISA were developed and applied to quantitative and qualitative detection of morphine. Under the optimal conditions for plate-based FLISA, the linear range spanned from 3.2 × 10 to 1 mg/L (R = 0.9905), and the detection limit was 2.7 × 10 mg/L. The visual detection limit for morphine by membrane-based flow-through FLISA was 0.01 mg/L. These results demonstrated that the developed fluorescence immunoassays could be applied as highly sensitive and convenient tools for rapid detection of morphine, which make it ideally suited for on-site screening of poppy shell added illegally in hot pot soup base.

摘要

开发了用于检测吗啡的量子点(QDs)标记抗体荧光免疫分析(FLISA)。使用含有羧基的量子点(CdSe/ZnS),通过作为偶联试剂的1-乙基-3-(3-二甲基氨基丙基)碳二亚胺盐酸盐/N-羟基琥珀酰亚胺来标记抗吗啡抗体。通过荧光光谱和凝胶电泳对CdSe/ZnS量子点标记的抗吗啡抗体(量子点标记的Ab)进行了表征。开发了基于平板的FLISA和基于硝酸纤维素膜的流通式FLISA,并将其应用于吗啡的定量和定性检测。在基于平板的FLISA的最佳条件下,线性范围为3.2×10至1 mg/L(R = 0.9905),检测限为2.7×10 mg/L。基于膜的流通式FLISA对吗啡的目视检测限为0.01 mg/L。这些结果表明,所开发的荧光免疫分析可作为高灵敏度且方便的工具用于快速检测吗啡,这使其非常适合现场筛查火锅汤底中非法添加的罂粟壳。

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