A serological aptamer-assisted proximity ligation assay for COVID-19 diagnosis and seeking neutralizing aptamers.
A serological aptamer-assisted proximity ligation assay for COVID-19 diagnosis and seeking neutralizing aptamers.
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DOI：
10.1039/d0sc03920a
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发表时间：
2020-10-12
期刊：
影响因子：
8.4
通讯作者：
Zhang J
中科院分区：
文献类型：
--
作者：
Liu R;He L;Hu Y;Luo Z;Zhang J
快速准确诊断COVID-19在当前疫情防控中发挥着至关重要的作用。尽管核酸和抗体检测前景看好，但要减少误诊，尤其是对无症状感染者的误诊，仍面临巨大挑战。在这里，我们报告了一种基于适体辅助邻近连接试验的高度特异性和超灵敏度检测血清COVID-19相关抗原的可推广方法。该传感器基于将两个适体探针结合到相同的蛋白质靶标上，使连接DNA区域非常接近，从而启动连接依赖性qPCR扩增。使用该系统，通过使用简单、均质和快速的检测工作流程，在202小时内将蛋白质识别转化为可检测的qPCR信号，定量检测血清核衣壳蛋白。此外，该系统还被转化为测量spike S1与其受体ACE 2之间特异性相互作用的通用平台，更重要的是，该系统证明了筛选和研究潜在中和适体的可行性。由于体外选择可以获得对许多COVID-19相关抗原具有选择性的适体，因此本文展示的方法将作为COVID-19诊断和治疗的重要工具。一种通用的适体辅助邻近连接系统改善了COVID-19的诊断，并允许评估潜在的中和适体。
Rapid and accurate diagnosis of COVID-19 plays an essential role in the current epidemic prevention and control. Despite the promise of nucleic acid and antibody tests, there is still a great challenge to reduce the misdiagnosis, especially for asymptomatic individuals. Here we report a generalizable method for highly specific and ultrasensitive detection of serum COVID-19-associated antigens based on an aptamer-assisted proximity ligation assay. The sensor is based on binding two aptamer probes to the same protein target that brings the ligation DNA region into close proximity, thereby initiating ligation-dependent qPCR amplification. Using this system, serum nucleocapsid protein has been detected quantitatively by converting protein recognition into a detectable qPCR signal using a simple, homogeneous and fast detection workflow in ∼2 hours. In addition, this system has also been transformed into a universal platform for measuring specific interactions between spike S1 and its receptor ACE2, and more importantly demonstrated the feasibility for screening and investigation of potential neutralizing aptamers. Since in vitro selection can obtain aptamers selective for many COVID-19-associated antigens, the method demonstrated here will serve as an important tool for the diagnosis and therapeutics of COVID-19. A versatile aptamer-assisted proximity ligation system improves diagnosis of COVID-19, and allows the evaluation of potential neutralizing aptamers.
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