Developing a Paper-Based Antigen Assay to Differentiate between Coronaviruses and SARS-CoV-2 Spike Variants
Developing a Paper-Based Antigen Assay to Differentiate between Coronaviruses and SARS-CoV-2 Spike Variants
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DOI：
10.1021/acs.analchem.0c05438
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发表时间：
2021-05-26
影响因子：
7.4
通讯作者：
Hamad-Schifferli, Kimberly
中科院分区：
文献类型：
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作者：
Hristov, Delyan;Rijal, Hom;Hamad-Schifferli, Kimberly
2019年12月，新冠肺炎首次出现在中国武汉。从那时起，它已成为一种全球流行病。强有力和可扩展的诊断战略对于遏制和监测大流行至关重要。RT-PCR是一种已知的、可靠的COVID-19诊断方法，可以区分SARS-CoV-2和其他病毒。然而，PCR依赖于位置，耗时且相对昂贵。因此，需要一种更灵活的方法，这种方法可以以现成的形式制作并更广泛地分发。基于纸张的免疫分析可以实现这一功能。在这里，我们介绍了迈向基于纸的测试的第一步，该测试可以区分各种冠状病毒（SARS-CoV-1、SARS-CoV-2和CoV-HKU1）的不同刺突蛋白，在一次检测中对HCoV-0C43和HCoV-229E的交叉反应性可以忽略不计，耗时不到30分钟。此外，我们的测试可以区分相同刺突蛋白的不同部分。这是通过一种改变的检测设计来完成的，有四个检测线位置，每个抗原建立一个独特的，可识别的结合模式。考虑了运行介质、免疫探针浓度和抗原干扰等因素的影响。我们发现，跑步介质对最终的结合模式有显著影响，其中人类唾液提供结果，而人类血清导致最低的信号质量。
COVID-19 first appeared in December of 2019 in Wuhan, China. Since then, it has become a global pandemic. A robust and scalable diagnostics strategy is crucial for containing and monitoring the pandemic. RT-PCR is a known, reliable method for COVID-19 diagnostics, which can differentiate between SARS-CoV-2 and other viruses. However, PCR is location-dependent, time-consuming, and relatively expensive. Thus, there is a need for a more flexible method, which may be produced in an off-the-shelf format and distributed more widely. Paper-based immunoassays can fulfill this function. Here, we present the first steps toward a paper-based test, which can differentiate between different spike proteins of various coronaviruses, SARS-CoV-1, SARS-CoV-2, and CoV-HKU1, with negligible cross-reactivity for HCoV-0C43 and HCoV-229E in a single assay, which takes less than 30 min. Furthermore, our test can distinguish between fractions of the same spike protein. This is done by an altered assay design with four test line locations where each antigen builds a unique, identifiable binding pattern. The effect of several factors, such as running media, immunoprobe concentration, and antigen interference, is considered. We find that running media has a significant effect on the final binding pattern where human saliva provides results while human serum leads to the lowest signal quality.