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首页> 美国卫生研究院文献>Pathogens >Application of DNA Aptamers and Quantum Dots to Lateral Flow Test Strips for Detection of Foodborne Pathogens with Improved Sensitivity versus Colloidal Gold
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Application of DNA Aptamers and Quantum Dots to Lateral Flow Test Strips for Detection of Foodborne Pathogens with Improved Sensitivity versus Colloidal Gold

机译：DNA适体和量子点在侧向流动测试条上的应用以检测与胶体金相比具有更高灵敏度的食源性病原体

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摘要

Preliminary studies aimed at improving the sensitivity of foodborne pathogen detection via lateral flow (LF) test strips by use of high affinity DNA aptamers for capture and reporter functions when coupled to red-emitting quantum dots (Qdot 655) are reported. A variety of DNA aptamers developed against Escherichia coli, Listeria monocytogenes, and Salmonella enterica were paired in capture and reporter combinations to determine which yielded the strongest detection of their cognate bacteria using a colloidal gold screening system. Several promising sandwich combinations were identified for each of the three bacterial LF strip systems. The best E. coli aptamer-LF system was further studied and yielded a visible limit of detection (LOD) of ~3,000 E. coli 8739 and ~6,000 E. coli O157:H7 in buffer. These LODs were reduced to ~300–600 bacterial cells per test respectively by switching to a Qdot 655 aptamer-LF system. Novel aspects of these assays such as the use of high levels of detergents to avoid quantum dot agglutination and enhance migration in analytical membranes, identification of optimal analytical membrane types, UV-immobilization of capture aptamers, and novel dual biotin/digoxigenin-end labeled aptamer streptavidin-colloidal gold or -Qdot 655 conjugates plus anti-digoxigenin antibody control lines are also discussed. In general, this work provides proof-of-principle for highly sensitive aptamer-Qdot LF strip assays for rapid foodborne pathogen detection.
机译：据报道，初步研究旨在通过使用高亲和力DNA适体与红色发射量子点（Qdot 655）耦合时的捕获和报告功能，通过侧流（LF）试纸来提高食源性病原体检测的灵敏度。将多种针对大肠杆菌，单核细胞增生李斯特菌和肠炎沙门氏菌的DNA适体在捕获和报告基因组合中配对，以确定使用胶体金筛选系统对它们的同源细菌的检测最强。对于三个细菌LF条带系统中的每一个，都确定了几种有希望的三明治组合。进一步研究了最佳的大肠杆菌适体-LF系统，并在缓冲液中产生了约3,000个大肠杆菌8739和6,000个O157：H7大肠杆菌的可见光检测极限（LOD）。通过切换到Qdot 655 aptamer-LF系统，每个测试将这些LOD分别减少到约300–600个细菌细胞。这些测定法的新颖性，例如使用高含量的去污剂以避免量子点凝集并增强分析膜中的迁移，鉴定最佳分析膜类型，将捕获适体进行紫外固定化，以及新型生物素/洋地黄毒苷双末端标记适体还讨论了抗生蛋白链菌素-胶体金或-Qdot 655偶联物以及抗地高辛配基的抗体对照品系。总的来说，这项工作为高灵敏度的适体-Qdot LF试纸条提供了原理证明，可快速检测食源性病原体。

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