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基于抗体共价固定“端到端”取向的侧向流条的检测评估。

The Evaluation of a Lateral Flow Strip Based on the Covalently Fixed "End-On" Orientation of an Antibody for Detection.

机构信息

Centre for Research and Development of Medical Diagnostic Laboratories (CMDL), Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand.

Department of Medical Technology, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand.

出版信息

Anal Chem. 2024 May 28;96(21):8543-8551. doi: 10.1021/acs.analchem.4c00533. Epub 2024 May 15.

DOI:10.1021/acs.analchem.4c00533
PMID:38748432
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11140673/
Abstract

In this study, the covalently fixed "end-on" orientation of a monoclonal antibody (mAb-) to amino terminated oligo (ethylene glycol)-capped gold nanoparticles (NH-TEG-AuNPs) was used to fabricate an in-house lateral flow strip (LFS), namely, the fixed "end-on" -mAb-NH-TEG-AuNPs LFS. The aim was to evaluate the performance of the fixed "end-on" -mAb-NH-TEG-AuNPs LFS in detecting . The proposed LFS enabled the sensitive detection of in 15 min with a visual limit of detection of 10 CFU/mL. Quantitative analysis indicated an LOD at 10 CFU/mL. The fixed "end-on" -mAb-NH-TEG-AuNPs LFS showed no cross-reactivity with other pathogenic bacteria and practical performance across different food matrices, including human blood, milk, and mushroom samples. Furthermore, the clinical performance of the fixed "end-on" -mAb-NH-TEG-AuNPs LFS for detecting was evaluated by using 12 clinical samples validated by the hemoculture method. It demonstrated excellent concordance with the reference methods, with no false-positive or false-negative results observed. Therefore, the fixed "end-on" -mAb-NH-TEG-AuNPs LFS serves as a promising candidate for a point-of-care test (POCT), enabling the rapid, precise, and highly sensitive detection of in clinical samples and contaminated food.

摘要

在这项研究中，利用单克隆抗体（mAb）与氨基封端的聚（乙二醇）封端的金纳米粒子（NH-TEG-AuNPs）的共价固定“端到端”取向，制备了一种内部横向流条（LFS），即固定“端到端”-mAb-NH-TEG-AuNPs LFS。目的是评估固定“端到端”-mAb-NH-TEG-AuNPs LFS 在检测中的性能。所提出的 LFS 能够在 15 分钟内实现对 的敏感检测，其视觉检测限为 10 CFU/mL。定量分析表明 LOD 为 10 CFU/mL。固定“端到端”-mAb-NH-TEG-AuNPs LFS 与其他病原菌无交叉反应，在不同食品基质（包括人血、牛奶和蘑菇样本）中具有实用性能。此外，通过使用经血液培养法验证的 12 个临床样本评估了固定“端到端”-mAb-NH-TEG-AuNPs LFS 检测 的临床性能。它与参考方法具有出色的一致性，没有观察到假阳性或假阴性结果。因此，固定“端到端”-mAb-NH-TEG-AuNPs LFS 可作为一种有前途的即时检测（POCT）候选物，能够在临床样本和污染食品中快速、精确和高度敏感地检测 。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58ec/11140673/f450a2c611ab/ac4c00533_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58ec/11140673/304c2e1a47b6/ac4c00533_0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58ec/11140673/f930c4049b6b/ac4c00533_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58ec/11140673/786e315ac97c/ac4c00533_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58ec/11140673/4ec6d98c6b0b/ac4c00533_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58ec/11140673/47ede4ffd80d/ac4c00533_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58ec/11140673/f450a2c611ab/ac4c00533_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58ec/11140673/304c2e1a47b6/ac4c00533_0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58ec/11140673/f930c4049b6b/ac4c00533_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58ec/11140673/786e315ac97c/ac4c00533_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58ec/11140673/4ec6d98c6b0b/ac4c00533_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58ec/11140673/47ede4ffd80d/ac4c00533_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58ec/11140673/f450a2c611ab/ac4c00533_0005.jpg

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