适体
指数富集配体系统进化
化学
严重急性呼吸综合征冠状病毒2型(SARS-CoV-2)
2019年冠状病毒病(COVID-19)
计算生物学
唾液
分子生物学
色谱法
生物
生物化学
医学
核糖核酸
基因
病理
传染病(医学专业)
疾病
作者
Rudi Liu,Jiuxing Li,Jim Gu,Bruno J. Salena,Yingfu Li
标识
DOI:10.1002/anie.202407049
摘要
Many aptamers have been generated by SELEX to recognize spike proteins of SARS‐CoV‐2, some of which have been engineered into dimeric and trimeric versions for enhanced affinity for diagnostic applications. However, no studies have been conducted to compare the utilities of monomeric, dimeric and trimeric aptamers in diagnostic assays with real clinical samples to answer the question of what levels of affinity an aptamer must have for accurate clinical diagnostics. Herein, we carried out a comparative study with two monomeric aptamers MSA1 and MSA5, one dimeric aptamer and two homotrimeric aptamers constructed with MSA1 and MSA5, with affinity varying by 1000‐fold. Using a colorimetric sandwich assay to analyze 48 human saliva samples, we found that the trimeric aptamer assay (Kd = ~10 pM) can identify the SARS‐CoV‐2 infection much more accurately than the dimeric aptamer assay (Kd = ~100 pM) and monomeric aptamer assay (Kd = ~10,000 pM). Based on the experimental data, we theoretically predict the levels of affinity an aptamer needs to possess to achieve 80‐100% sensitivity and 100% specificity. The findings from this study highlight the need for deriving very high affinity aptamers to enable highly accurate detection of viral infection for future pandemics.
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