Proof-of-concept for speedy development of rapid and simple at-home method for potential diagnosis of early COVID-19 mutant infections using nanogold and aptamer.
Proof-of-concept for speedy development of rapid and simple at-home method for potential diagnosis of early COVID-19 mutant infections using nanogold and aptamer.
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DOI：
10.1016/j.nano.2022.102590
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发表时间：
2022-09
影响因子：
5.4
通讯作者：
Guo, Peixuan
中科院分区：
文献类型：
--
作者：
Ellipilli, Satheesh;Wang, Hongzhi;Lee, Wen-Jui;Shu, Dan;Guo, Peixuan
新冠肺炎基因的单链阳性性质导致其基因组鉴定的校对效率较低。因此，新冠肺炎突变株一直在快速进化。除了Alpha、Beta、Gamma、Delta和Omicron变种外，目前还在流传Omicron的亚变种，包括BA.4、BA.5和BA.2.12.1。因此，快速发展一种快速、简便、易行的诊断方法来应对新的变异冠状病毒感染是至关重要的。目前已建立了多种新冠肺炎的检测方法，如逆转录聚合酶链式反应和抗体检测等。然而，前者费时、费力且昂贵，而后者依赖于抗体的产生，不适合病毒感染的早期诊断。有许多侧向流动方法可用，但可能不适合检测突变，在这里，我们证明了通过结合一种新的适体，快速开发一种简单、快速和经济高效的突变新冠肺炎感染的家庭诊断方法的概念。这个想法是使用目前市场上可用的横向流动新冠肺炎诊断系统，或者使用一种现有的抗体用于横向流动硝化纤维素滤器。为了验证这一概念，将针对SPAKE蛋白(S蛋白)的DNA适配子连接到纳米金颗粒上，作为检测探针。一种针对COVID病毒颗粒上过度表达的S蛋白的抗体被用作固定在硝酸纤维素膜上的第二个探针。将适体结合的纳米颗粒与Spike蛋白孵育半小时，并测试它们与锚定在硝酸纤维素膜上的抗体的结合能力。由于抗原(S蛋白)与适配子和抗体之间的相互作用，金纳米颗粒被显示在硝酸纤维素膜上。因此，可以在2小时内获得病毒抗原的检测，而诊断试剂的成本不到5美元。未来，只要报道新出现的病毒表面蛋白的突变，就可以在几天内通过肽合成或基因克隆产生与突变相对应的多肽或蛋白质。RNA或DNA适配子可以通过SELEX快速生成。一种针对尖峰蛋白(S蛋白)的金标适体将作为检测探针。市场上已有的任何带有固定化抗体的侧向流动诊断试剂盒，或者仅仅是结合新冠肺炎病毒的抗体，都可以用作固定在硝酸纤维素上的第二个探针。如果临床试验验证了该方法的可行性和特异性，则患者可以在家中进行该诊断方法。
The positive single-stranded nature of COVID-19 mRNA led to the low proof-reading efficacy for its genome authentication. Thus mutant covid-19 strains have been rapidly evolving. Besides Alpha, Beta, Gamma, Delta, and Omicron variants, currently, subvariants of omicron are circulating, including BA.4, BA.5, and BA.2.12.1. Therefore, the speedy development of a rapid, simple, and easier diagnosis method to deal with new mutant covid viral infection is critically important. Many diagnosis methods have been developed for COVID-19 detection such as RT-PCR and antibodies detection. However, the former is time-consuming, laborious, and expensive, and the latter relies on the production of antibodies making it not suitable for the early diagnosis of viral infection. Many lateral-flow methods are available but might not be suitable for detecting the mutants, Here we proved the concept for the speedy development of a simple, rapid, and cost-effective early at-home diagnosis method for mutant Covid-19 infection by combining a new aptamer. The idea is to use the current lateral flow Covid-19 diagnosis system available in the market or to use one existing antibody for the Lateral Flow Nitrocellulose filter. To prove the concept, the DNA aptamer specific to spike proteins (S-proteins) was conjugated to gold nanoparticles and served as a detection probe. An antibody that is specific to spike proteins overexpressed on COVID viral particles was used as a second probe immobilized to the nitrocellulose membrane. The aptamer conjugated nanoparticles were incubated with spike proteins for half an hour and tested for their ability to bind to antibodies anchored on the nitrocellulose membrane. The gold nanoparticles were visualized on the nitrocellulose membrane due to interaction between the antigen (S-protein) with both the aptamer and the antibody. Thus, the detection of viral antigen can be obtained within 2 h, with a cost of less than $5 for the diagnosis reagent. In the future, as long as the mutant of the newly emerged viral surface protein is reported, a peptide or protein corresponding to the mutation can be produced by peptide synthesis or gene cloning within several days. An RNA or DNA aptamer can be generated quickly via SELEX. A gold-labeled aptamer specific to spike proteins (S-proteins) will serve as a detection probe. Any available lateral-flow diagnosis kits with an immobilized antibody that has been available on the market, or simply an antibody that binds COVID-19 virus might be used as a second probe immobilized on the nitrocellulose. The diagnosis method can be carried out by patients at home if a clinical trial verifies the feasibility and specificity of this method.
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