PDF(Pubmed)
Abstract：
The emergence of the SARS-CoV-2 virus, an unknown strain of coronavirus, has resulted in severe acute respiratory syndrome with high mortality rates worldwide. Due to the possibility of asymptomatic carriers, late diagnosis of infected individuals can lead to uncontrollable transmission of the disease, making early and accurate detection crucial in controlling the spread of the virus. In this study we identified high-binding-affinity aptamers targeting various strains of the SARS-CoV2 (COVID-19) virus, using the GO-Cell-SELEX (Graphene Oxide- Systematic Evolution of Ligands by Exponential Enrichment) strategy. A total of 96 aptamers were developed through 11 rounds of GO-Cell-SELEX from a random 40 nucleotide single-strand DNA (ssDNA) aptamer library. Using the surface plasmon resonance (SPR) method, the dissociation constant (Kd) values of all aptamers were calculated and two aptamers 52 and 91 with Kd 50 and 61 were selected for enzyme-linked apta-sorbent assay (ELASA). Aptamer 91 could detect various strains of the virus in above 97% of clinical samples obtained from nasopharyngeal swaps (NPS) specimens kept in viral transport media (VTM), confirmed by real-time PCR assay at COVID-19 Reference Diagnostic Laboratory of Iran, Pasture Institute. Aptamer 52 could detect the SARS-CoV2 virus in a competitive lateral flow assay (LFA) to be considered for a future designed kit. These two simple, specific, and sensitive tests can be used in combination for rapid and early diagnosis of various strains of the COVID-19 virus. Our results suggest that these two discovered aptamers present an opportunity for developing a new rapid aptamer-based coronavirus diagnostic kit.
摘要：
SARS-CoV-2病毒的出现,一种未知的冠状病毒，导致了严重的急性呼吸系统综合症，全球死亡率很高。由于无症状携带者的可能性，对感染者的晚期诊断可能导致疾病的无法控制的传播，使早期和准确的检测对于控制病毒传播至关重要。在这项研究中，我们确定了针对SARS-CoV2（COVID-19）病毒各种毒株的高结合亲和力适体，使用GO-Cell-SELEX(石墨烯氧化物-指数富集配体的系统进化)策略。通过11轮GO-Cell-SELEX从随机40个核苷酸的单链DNA(ssDNA)适体文库开发了总共96个适体。使用表面等离子体共振（SPR）方法，计算了所有适体的解离常数(Kd)值,并选择了Kd为50和61的两个适体52和91用于酶联apta吸附测定(ELASA).适体91可以在保存在病毒转运培养基（VTM）中的鼻咽交换（NPS）标本中获得的超过97％的临床样品中检测到各种病毒株，通过伊朗COVID-19参考诊断实验室的实时PCR检测证实，牧场研究所。适体52可以在竞争性侧流测定（LFA）中检测SARS-CoV2病毒，以考虑将来设计的试剂盒。这两个简单的，具体,和敏感测试可结合使用，以快速和早期诊断各种株的COVID-19病毒。我们的结果表明，这两个发现的适体为开发新的快速基于适体的冠状病毒诊断试剂盒提供了机会。
