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基于亲和素-生物素复合物的侧向流动分析联合等温扩增技术对猪圆环病毒 2 的高灵敏快速检测。

Highly sensitive and rapid detection of porcine circovirus 2 by avidin-biotin complex based lateral flow assay coupled to isothermal amplification.

机构信息

Department of Bionanotechnology, Gachon University, Seongnam, 461-70, Republic of Korea.

R&D Center, Philmedi Ltd, Seongnam, 461-70, Republic of Korea.

出版信息

Anal Methods. 2021 Oct 8;13(38):4429-4436. doi: 10.1039/d1ay01189h.

Abstract

In this study, a new platform for the detection of porcine circovirus 2 was developed by avidin-biotin complex based lateral flow assay (LAMP-LFA). Improved detection sensitivity was attained by using loop mediated isothermal amplification (LAMP) with a low limit of detection (LOD), so the platform can be used to detect even early or asymptomatic stages of infection. LFA, which requires no specialized equipment, facilitates the use of point-of-care (POC) tests. Therefore, by applying LFA, the result can be confirmed accurately with the naked eye. Moreover, this platform has a unique structure using a single-tag detection system. The avidin-biotin interaction is the strongest interaction between proteins and has a higher value than antigen-antibody interactions. Thus, the results are stable and can be clearly confirmed. The high sensitivity of LAMP-LFA enables all steps to be completed in 30 min. As a result, it could be applied to different targets, such as other pathogens. Future POC diagnostic studies are expected to be of great practical benefit.

摘要

本研究基于亲和素-生物素复合物建立了一种新的猪圆环病毒 2 检测平台,即侧流免疫层析检测-环介导等温扩增法(LAMP-LFA)。通过低检测限(LOD)的环介导等温扩增(LAMP)提高了检测灵敏度,因此该平台可用于检测感染的早期或无症状阶段。不需要特殊设备的 LFA 便于进行即时检测(POC)测试。因此,通过应用 LFA,可以用肉眼准确地确认结果。此外,该平台采用独特的单标记检测系统结构。亲和素-生物素相互作用是蛋白质之间最强的相互作用,其解离常数(Kd)值比抗原-抗体相互作用更小。因此,结果稳定且可以清晰地确认。LAMP-LFA 的高灵敏度使得所有步骤都可以在 30 分钟内完成。因此,它可以应用于不同的靶标,如其他病原体。未来的即时检测诊断研究有望具有重要的实际应用价值。

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