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基于适体功能化纳米金颗粒染料的微污染物筛查侧向流动免疫层析试纸条的研制,第二部分:黄曲霉毒素B1和氯霉素的实验验证

Development of Lateral Flow Immunochromatographic Strips for Micropollutant Screening Using Colorants of Aptamer-Functionalized Nanogold Particles, Part II: Experimental Verification with Aflatoxin B1 and Chloramphenicol.

作者信息

Zhang Shan, Zhao Shuai, Wang Sai, Liu Jiahui, Dong Yiyang

机构信息

Beijing University of Chemical Technology, College of Life Science and Technology, Beijing 100029, People's Republic of China.

Ocean University of China, College of Food Science and Engineering, Lab of Nucleic Acid Chemistry and Biotechnology, Qingdao 266003, China.

出版信息

J AOAC Int. 2018 Sep 1;101(5):1408-1414. doi: 10.5740/jaoacint.18-0056. Epub 2018 May 9.

Abstract

Lateral flow immunochromatographic strips based on colorants of aptamer-functionalized nanogold particles were developed for the detection of micropollutants aflatoxin B1 (AFB1) and chloramphenicol (CAP). The lateral flow immunochromatographic strip was based on a competitive reaction of thiolated-aptamer between micropollutants and bio-DNA probe-streptavidin as capture material immobilized at the test line. General crucial parameters that might influence the sensitivity have been systematically investigated. To test the effectiveness and applicability of the optimized conditions, two structurally unrelated micropollutants, that is, AFB1 and CAP, were chosen for detection. In the present study, lateral flow immunochromatographic strips for AFB1 and CAP analysis by combining the high selectivity and affinity of aptamers with the unique optical properties of nanogold in municipal water samples were reported for the first time. With the optimized conditions, the immunochromatographic strip showed a visual LOD of 10 ppb and a quantitative LOD of 1.05 ppb using an immunochromatographic reader for AFB1 detection and a quantitative LOD of 63.4 ppb using an immunochromatographic reader for CAP detection. Furthermore, the sensitive strip provided a good linear detection range of approximately 0-50 ppm for AFB1 detection and a wider liner detection range of approximately 0-160 ppm for CAP detection. Moreover, the immunochromatographic strip provided recovery rates for water samples of 90-110% in the AFB1 analysis and 84-108% in the CAP analysis. The results demonstrated that the immunochromatographic strip has excellent potential for wide applicability and verified that the strip methods for the optimized conditions are applicable to a variety of micropollutants. The lateral flow immunochromatographic strip could be used as a simple, rapid, and efficient screening tool for rapid on-site detection of a variety of micropollutants.

摘要

基于适体功能化纳米金颗粒着色剂的侧流免疫层析试纸条被开发用于检测微污染物黄曲霉毒素B1(AFB1)和氯霉素(CAP)。侧流免疫层析试纸条基于微污染物与生物DNA探针 - 链霉亲和素之间硫醇化适体的竞争反应,生物DNA探针 - 链霉亲和素作为捕获材料固定在检测线上。系统地研究了可能影响灵敏度的一般关键参数。为了测试优化条件的有效性和适用性,选择了两种结构不相关的微污染物,即AFB1和CAP进行检测。在本研究中,首次报道了通过将适体的高选择性和亲和力与纳米金在城市水样中的独特光学性质相结合,用于AFB1和CAP分析的侧流免疫层析试纸条。在优化条件下,免疫层析试纸条对AFB1检测使用免疫层析读数器时,视觉检测限为10 ppb,定量检测限为1.05 ppb;对CAP检测使用免疫层析读数器时,定量检测限为63.4 ppb。此外,该灵敏试纸条对AFB1检测提供了约0 - 50 ppm的良好线性检测范围,对CAP检测提供了约0 - 160 ppm的更宽线性检测范围。而且,免疫层析试纸条在AFB1分析中对水样的回收率为90 - 110%,在CAP分析中为84 - 108%。结果表明,免疫层析试纸条具有广泛应用的优异潜力,并验证了优化条件下的试纸条方法适用于多种微污染物。侧流免疫层析试纸条可作为一种简单、快速且高效的筛选工具,用于快速现场检测多种微污染物。

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