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一种用于快速筛查乙型流感病毒的基于荧光的简易侧向流动检测平台。

A simple fluorescence-based lateral flow test platform for rapid influenza B virus screening.

作者信息

Wiriyachaiporn Natpapas, Sirikaew Siriwan, Bamrungsap Suwussa, Limcharoen Thanchanok, Polkankosit Pannarai, Roeksrungruang Pimporn, Ponlamuangdee Kanyawan

机构信息

National Nanotechnology Center (NANOTEC), National Science and Technology Development Agency (NSTDA), Pathumthani 12120, Thailand.

出版信息

Anal Methods. 2021 Apr 14;13(14):1687-1694. doi: 10.1039/d0ay01988g. Epub 2021 Mar 22.

Abstract

A simple fluorescence-based lateral flow test platform for rapid influenza B virus screening as a model target molecule was successfully developed. In this work, Cy5-loaded silica nanoparticles were directly conjugated to monoclonal antibodies, specific to the influenza B nucleoprotein, via a direct physisorption method and used as detector probes. Using this approach, the signal response to the detection was further determined using a fluorescent signal intensity measurement method via a portable reader, in combination with fluorescence imaging analysis. The degree to which the fluorescence signal response is detected is proportional to the amount of the target virus protein present in the system, reflected by the accumulation of the formed particle-antibody conjugates within the test system. Under optimized conditions, the system is capable of detecting the influenza B virus protein at a level of 0.55 μg per test within 30 min, using small sample volumes as low as 100 μL (R = 0.9544). In addition to its simplicity, further application of the system in detecting the influenza B virus protein was demonstrated using the viral transport media as specimen matrices. It was also shown that the system can perform the detection without cross-reactivity to other closely related respiratory viruses.

摘要

成功开发了一种基于荧光的简单侧向流动检测平台,用于快速筛查乙型流感病毒作为模型目标分子。在这项工作中,通过直接物理吸附法将负载Cy5的二氧化硅纳米颗粒直接与针对乙型流感核蛋白的单克隆抗体偶联,并用作检测探针。使用这种方法,通过便携式读数器结合荧光成像分析,使用荧光信号强度测量方法进一步确定对检测的信号响应。检测到的荧光信号响应程度与系统中存在的目标病毒蛋白量成正比,这通过测试系统中形成的颗粒-抗体缀合物的积累来反映。在优化条件下,该系统能够在30分钟内使用低至100μL的小样本体积检测乙型流感病毒蛋白,检测水平为每次测试0.55μg(R = 0.9544)。除了其简单性之外,还使用病毒运输介质作为样本基质证明了该系统在检测乙型流感病毒蛋白方面的进一步应用。还表明该系统可以进行检测,而不会与其他密切相关的呼吸道病毒发生交叉反应。

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