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基于骆驼单域抗体的苏云金芽胞杆菌 Cry1B 毒素的灵敏检测。

Sensitive detection of Bacillus thuringiensis Cry1B toxin based on camel single-domain antibodies.

机构信息

CAS Key Laboratory of Receptor Research, Shanghai Institute of Material Medical, Chinese Academy of Sciences, Shanghai, China.

University of Chinese Academy of Sciences, Beijing, China.

出版信息

Microbiologyopen. 2018 Aug;7(4):e00581. doi: 10.1002/mbo3.581. Epub 2018 Feb 24.

Abstract

Bt Cry1B toxin, a residue in insect-resistant transgenic plants, has been identified to be harmful to human health. Therefore, it is urgent to detect the Cry1B toxin level in each kind of transgenic plant. Nbs, with prominently unique physiochemical properties, are becoming more and more promising tools in the detection of target antigens. In this study, an immune phage display library that was of high quality was successfully constructed for the screening of Cry1B-specific Nbs with excellent specificity, affinity, and thermostable. Subsequently, a novel sandwich ELISA for Cry1B detection was established, which was based on the biotin-streptavidin system using these aforementioned Nbs. This established detection system presented a linear working range from 5 to 1000 ng ml and a low detection limit of 3.46 ng ml . The recoveries from spiked samples were in the range of 82.51%-113.56% with a relative standard deviation (RSD) lower than 5.00%. Taken together, the proposed sandwich ELISA would be a potential method for the detection of Cry1B toxin in transgenic Bt plants specifically and sensitively.

摘要

Bt Cry1B 毒素是抗虫转基因植物中的一种残留物质,已被确定对人类健康有害。因此,迫切需要检测各种转基因植物中的 Cry1B 毒素水平。Nbs 具有独特的物理化学性质,作为目标抗原的检测工具越来越有前途。本研究成功构建了一种高质量的免疫噬菌体展示文库,用于筛选具有优异特异性、亲和力和热稳定性的 Cry1B 特异性 Nbs。随后,建立了一种基于生物素-链霉亲和素系统的新型 Cry1B 检测夹心 ELISA。该检测系统的线性工作范围为 5 至 1000ng/ml,检测限低至 3.46ng/ml。添加样品的回收率在 82.51%至 113.56%之间,相对标准偏差(RSD)低于 5.00%。总之,该夹心 ELISA 有望成为一种特异性和灵敏性检测转基因 Bt 植物中 Cry1B 毒素的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac06/6079177/290ee62f7558/MBO3-7-e00581-g001.jpg

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