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基于免疫磁珠的生物素-链霉亲和素系统用于高效检测农产品中的黄曲霉毒素B。

Immunomagnetic bead-based biotin-streptavidin system for highly efficient detection of aflatoxin B in agricultural products.

作者信息

Li Ming, Zhang Yuanyuan, Zhao Rujin, Liu Zhenjiang, Hong Xia, Cui Yin, Xue Yonglai, Du Daolin

机构信息

Institute of Environment and Ecology, Institute of Environmental Health and Ecological Security, School of the Environment and Safety Engineering, Jiangsu University Zhenjiang 212013 P. R. China

Department of Environmental Engineering, School of the Environment and Safety Engineering, Jiangsu University Zhenjiang 212013 P. R. China.

出版信息

RSC Adv. 2018 Jul 19;8(46):26029-26035. doi: 10.1039/c8ra04460k.

Abstract

The potential homogeneous assay employing immunomagnetic beads (IMB) has been receiving attention as a screening tool in food-safety control; the method is simple, efficient, and does not require long incubation times or complex separation steps. In this study, a homogeneous immunoassay has been successfully developed and applied in the determination of aflatoxin B (AFB) contamination in agricultural products by coupling IMB and the biotin-streptavidin (BSA) (BSA-IMB) system. Under optimal conditions, the limit of detection (LOD, IC), half-maximal inhibition concentration (IC) and detection range (IC-IC) of BSA-IMB are 0.00579, 0.573 and 0.0183-17.9 ng mL, respectively, for AFB. The detection of AFB by BSA-IMB can be achieved in 40 min (ELISA needs at least 180 min). The cross-reactivities of BSA-IMB with its analogues are negligible (<3.82%); these results indicate high selectivity. The spiked recoveries are in the range from 89.6 to 118.2% with relative standard deviations (RSDs) of 3.4 to 13.2% for AFB in agricultural product samples. Furthermore, the results of BSA-IMB for authentic samples show reliability and high correlation of 0.9928 with an HPLC-fluorescence detector. The proposed BSA-IMB system is demonstrated to be a satisfactory tool for homogeneous, efficient, sensitive, and alternative detection of AFB in a wide detection range for agricultural product samples.

摘要

采用免疫磁珠(IMB)的潜在均相分析方法作为食品安全控制中的一种筛选工具受到了关注;该方法简单、高效,不需要长时间孵育或复杂的分离步骤。在本研究中,通过将免疫磁珠与生物素-链霉亲和素(BSA)(BSA-IMB)系统相结合,成功开发了一种均相免疫分析方法,并将其应用于农产品中黄曲霉毒素B(AFB)污染的测定。在最佳条件下,BSA-IMB对AFB的检测限(LOD,IC)、半最大抑制浓度(IC)和检测范围(IC-IC)分别为0.00579、0.573和0.0183-17.9 ng/mL。BSA-IMB检测AFB可在40分钟内完成(酶联免疫吸附测定至少需要180分钟)。BSA-IMB与其类似物的交叉反应可忽略不计(<3.82%);这些结果表明其具有高选择性。在农产品样品中,AFB的加标回收率为89.6%至118.2%,相对标准偏差(RSD)为3.4%至13.2%。此外,真实样品的BSA-IMB检测结果显示出可靠性,与高效液相色谱-荧光检测器的相关性高达0.9928。所提出的BSA-IMB系统被证明是一种用于农产品样品中AFB的均相、高效、灵敏且可替代的检测工具,具有较宽的检测范围。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8452/9083124/bdbfbbc4e3a7/c8ra04460k-f1.jpg

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