Multi-chromatic and multi-component lateral flow immunoassay for simultaneous detection of CP4 EPSPS, Bt-Cry1Ab, Bt-Cry1Ac, and PAT/bar proteins in genetically modified crops

IF 5.3 2区化学 Q1 CHEMISTRY, ANALYTICAL

Microchimica Acta Pub Date : 2024-12-16 DOI:10.1007/s00604-024-06853-9

Yao Yang, Zini Zhang, Zhi Wang, Ruxin Pan, Huimin Wu, Shanshan Zhai, Gang Wu, Wei Fu, Hongfei Gao

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Abstract

A multi-chromatic and multi-component lateral flow immunoassay (MCMC-LFIA) was developed for simultaneous detection of CP4 EPSPS, Bt-Cry1Ab, Bt-Cry1Ac, and PAT/bar proteins in genetically modified (GM) crops. Captured antibodies specific to these exogenous proteins were separately immobilized on a nitrocellulose membrane as test zones. Multi-colored microspheres, used as visible multi-probes, were conjugated with corresponding antibodies and sprayed on the conjugate pad. The assay results can be visually interpreted within 10 min by observing the appearance of colored bands. The MCMC-LFIA demonstrated high sensitivity, with detection of limits of 7.8 ng/mL for CP4 EPSPS and 2.5 ng/mL for Bt-Cry1Ab, Bt-Cry1Ac, and PAT/bar proteins, significantly improving the performance of previously reported LFIAs. The MCMC-LFIA exhibited excellent specificity and was validated for practical use in field-based applications. The proposed MCMC-LFIA offers a rapid, sensitive, and user-friendly tool for the on-site large-scale screening of GM materials.

Graphical Abstract

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为同时检测转基因作物中的 CP4 EPSPS、Bt-Cry1Ab、Bt-Cry1Ac 和 PAT/bar 蛋白，开发了一种多色谱和多组分横向流动免疫分析法（MCMC-LFIA）。捕获的针对这些外源蛋白的特异性抗体被分别固定在硝酸纤维素膜上作为检测区。多色微球作为可见多探针，与相应的抗体共轭并喷洒在共轭垫上。通过观察彩色条带的出现，可在 10 分钟内直观地解读检测结果。MCMC-LFIA 具有很高的灵敏度，CP4 EPSPS 的检测限为 7.8 纳克/毫升，Bt-Cry1Ab、Bt-Cry1Ac 和 PAT/bar 蛋白的检测限为 2.5 纳克/毫升，大大提高了之前报道的 LFIA 的性能。MCMC-LFIA 具有出色的特异性，并通过了现场实际应用的验证。拟议的 MCMC-LFIA 为现场大规模筛选转基因材料提供了一种快速、灵敏和用户友好的工具。

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来源期刊

Microchimica Acta 化学-分析化学

CiteScore

9.80

自引率

5.30%

发文量

410

审稿时长

2.7 months

期刊介绍： As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.