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用于检测HIV-1表面糖蛋白的基于液晶的结合测定法。

Liquid Crystal Based Binding Assay for Detecting HIV-1 Surface Glycoprotein.

作者信息

Abbasi Amna Didar, Hussain Zakir, Liaqat Usman, Arif Dooa, Yang Kun-Lin

机构信息

Department of Materials Engineering, School of Chemical and Materials Engineering, National University of Sciences and Technology, Islamabad, Pakistan.

Department of Chemical and Biomolecular Engineering, National University of Singapore, Singapore, Singapore.

出版信息

Front Chem. 2021 Apr 26;9:668870. doi: 10.3389/fchem.2021.668870. eCollection 2021.

Abstract

Surface protein gp-120 of HIV-1 virus plays an important role in the infection of HIV-1, but detection of gp-120 during the early stage of infection is very difficult. Herein, we report a binding bioassay based on an RNA aptamer B40t77, which binds specifically to gp-120. The bioassay is built upon a hydrophobic glass slide with surface immobilized gp-120. When the glass surface is incubated in a solution containing B40t77, the aptamer is able to bind to gp-120 specifically and remove it from the surface after a short incubation time of 30 min. The result of the binding event can be amplified by using liquid crystal (LC) into optical signals in the final step. By using this bioassay, we are able to detect as low as 1 μg/ml of gp-120 with high specificity within 30 min. No response is obtained when gp-120 is replaced by other protein such as bovine serum albumin (BSA). This is the first qualitative bioassay which provides a simple way for the detection of gp-120 with the naked eye. The assay is robust, low-cost and does not require additional labeling. Thus, the bioassay is potentially useful for the early detection of HIV-1 in resources-limited regions.

摘要

HIV-1病毒的表面蛋白gp-120在HIV-1感染过程中起着重要作用,但在感染早期检测gp-120非常困难。在此,我们报道了一种基于RNA适配体B40t77的结合生物测定法,该适配体可特异性结合gp-120。该生物测定法构建在表面固定有gp-120的疏水载玻片上。当将玻璃表面在含有B40t77的溶液中孵育时,该适配体能够特异性结合gp-120,并在短时间孵育30分钟后将其从表面去除。结合事件的结果可在最后一步通过使用液晶(LC)放大为光学信号。通过使用这种生物测定法,我们能够在30分钟内以高特异性检测低至1μg/ml的gp-120。当用其他蛋白质如牛血清白蛋白(BSA)替代gp-120时,未获得响应。这是第一种定性生物测定法,为肉眼检测gp-120提供了一种简单方法。该测定法稳健、低成本且无需额外标记。因此,该生物测定法在资源有限地区对HIV-1的早期检测可能具有重要作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/47b4/8107394/ff69f6e26bee/fchem-09-668870-g0001.jpg

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