基于抗独特型纳米抗体-噬菌体展示的实时免疫-PCR 法用于灵敏、同时和定量检测谷物中的总黄曲霉毒素和玉米赤霉烯酮。

Anti-idiotypic nanobody-phage display-mediated real-time immuno-PCR for sensitive, simultaneous and quantitative detection of total aflatoxins and zearalenone in grains.

机构信息

Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs, Wuhan 430062, PR China.

Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture and Rural Affairs, Wuhan 430062, PR China.

出版信息

Food Chem. 2019 Nov 1;297:124912. doi: 10.1016/j.foodchem.2019.05.186. Epub 2019 May 28.

DOI:10.1016/j.foodchem.2019.05.186

PMID:31253263

Abstract

An anti-idiotypic nanobody-phage display-mediated immuno-polymerase chain reaction (PD-IPCR) method was developed for simultaneous quantitative detection of total aflatoxins and zearalenone in cereals. Two phages, displaying the variable domain of the heavy chain anti-idiotypic nanobody that binds aflatoxin- or zearalenone-specific monoclonal antibody (1C11 or 2D3), were used as competitors for corresponding analytes. Specific DNA sequences encoding anti-idiotypic nanobodies were used to design the primers for PCR amplification. The results indicated that detection limits for total aflatoxins and zearalenone in a sample were 0.03 and 0.09 ng mL, respectively. Recoveries of spiked aflatoxins and zearalenone were 80-118% and 76.7-111%, respectively. Validation results were in good agreement with the gold-standard high-performance liquid chromatography method. This report is the first to describe PD-IPCR for simultaneous quantitative detection of total aflatoxins and zearalenone in cereals.

摘要

建立了一种抗独特型纳米体-噬菌体展示介导的免疫聚合酶链反应（PD-IPCR）方法，用于同时定量检测谷物中的总黄曲霉毒素和玉米赤霉烯酮。两种噬菌体展示了与黄曲霉毒素或玉米赤霉烯酮特异性单克隆抗体（1C11 或 2D3）结合的重链抗独特型纳米体的可变结构域，用作相应分析物的竞争物。特异性编码抗独特型纳米体的 DNA 序列被用于设计 PCR 扩增的引物。结果表明，样品中总黄曲霉毒素和玉米赤霉烯酮的检测限分别为 0.03 和 0.09 ng mL。黄曲霉毒素和玉米赤霉烯酮的加标回收率分别为 80-118%和 76.7-111%。验证结果与金标准高效液相色谱法吻合良好。本报告首次描述了 PD-IPCR 用于同时定量检测谷物中的总黄曲霉毒素和玉米赤霉烯酮。