Quantitative Detection of the CP4-EPSPS Protein in Genetically Modified Soybeans Using a Novel Fluorescence Immunoassay Assay.

IF 1.9 2区 农林科学 Q3 FOOD SCIENCE & TECHNOLOGY Foodborne pathogens and disease Pub Date : 2024-11-01 Epub Date: 2024-08-07 DOI:10.1089/fpd.2024.0024
Xue Xu, Lu Jin, Xiaolei Zhu, Jiong Li
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Abstract

Protein-based detection methods, enzyme-linked immunosorbent assays (ELISAs) and lateral flow strips, have been widely used for rapid, specific, and sensitive detection of genetically modified organisms (GMOs). However, the traditional ELISA method for the quantitative detection of GMOs has certain limitations. Herein, a quantum dot (QD)-based fluorescence-linked immunosorbent assay was developed using QDs as fluorescent markers for the detection of glyphosate-resistant protein (CP4-EPSPS) in the MON89788 soybean. The end-point fluorescent detection system was carried out using QDs conjugated with a goat anti-rabbit secondary antibody. Compared with the conventional sandwich ELISA method, the newly developed fluorescence-linked immunosorbent assay was highly sensitive and accurate for detecting the CP4-EPSPS protein. The quantified linearity was achieved in the range of 0.05-5% (w/w) for the MON89788 soybean sample. The recovery of protein extracted from mixed MON89788 soybean samples ranged from 87.67% to 116.83%. The limits of detection and limits of quantification were 0.7101 and 2.152 pg/mL, respectively. All of the results indicated that the QD-based fluorescence-linked immunosorbent assay was a highly specific and sensitive method for monitoring the CP4-EPSPS protein in GMOs.

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利用新型荧光免疫测定法定量检测转基因大豆中的 CP4-EPSPS 蛋白。
基于蛋白质的检测方法--酶联免疫吸附测定法(ELISA)和侧向流动条,已被广泛用于快速、特异、灵敏地检测转基因生物(GMOs)。然而,传统的酶联免疫吸附法在定量检测转基因生物方面存在一定的局限性。本文以量子点(QD)为荧光标记,开发了一种基于量子点(QD)的荧光联免疫吸附测定法,用于检测 MON89788 大豆中的抗草甘膦蛋白(CP4-EPSPS)。端点荧光检测系统采用 QDs 与山羊抗兔二抗连接。与传统的夹心酶联免疫吸附法相比,新开发的荧光联免疫吸附法检测 CP4-EPSPS 蛋白的灵敏度和准确度都很高。对 MON89788 大豆样品的定量线性范围为 0.05-5%(w/w)。从混合的 MON89788 大豆样品中提取的蛋白质的回收率为 87.67% 至 116.83%。检测限和定量限分别为 0.7101 和 2.152 pg/mL。所有结果表明,基于 QD 的荧光连接免疫吸附分析法是一种特异性和灵敏度极高的监测转基因生物中 CP4-EPSPS 蛋白的方法。
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来源期刊
Foodborne pathogens and disease
Foodborne pathogens and disease 医学-食品科技
CiteScore
5.30
自引率
3.60%
发文量
80
审稿时长
1 months
期刊介绍: Foodborne Pathogens and Disease is one of the most inclusive scientific publications on the many disciplines that contribute to food safety. Spanning an array of issues from "farm-to-fork," the Journal bridges the gap between science and policy to reduce the burden of foodborne illness worldwide. Foodborne Pathogens and Disease coverage includes: Agroterrorism Safety of organically grown and genetically modified foods Emerging pathogens Emergence of drug resistance Methods and technology for rapid and accurate detection Strategies to destroy or control foodborne pathogens Novel strategies for the prevention and control of plant and animal diseases that impact food safety Biosecurity issues and the implications of new regulatory guidelines Impact of changing lifestyles and consumer demands on food safety.
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