基因分型
计算生物学
环介导等温扩增
数字聚合酶链反应
脱氧核酶
分析物
病毒学
2019年冠状病毒病(COVID-19)
核酸
检出限
核糖核酸
纳斯巴
生物
DNA
传染病(医学专业)
聚合酶链反应
化学
遗传学
医学
基因
基因型
色谱法
疾病
病理
作者
Kefan Yang,John C. Chaput
标识
DOI:10.1021/acssynbio.3c00105
摘要
The COVID-19 pandemic, caused by the SARS-CoV-2 virus, exposed a pressing need for new public health tools for pathogen detection, disease diagnosis, and viral genotyping. REVEALR (RNA-encoded viral nucleic acid analyte reporter) is an isothermal DNAzyme-based point-of-care diagnostic that functions with a detection limit of ∼10 copies/μL when coupled with a preamplification step and can be utilized for viral genotyping of SARS-CoV-2 variants of concern through base pair mismatch recognition in a competitive binding format. Here, we describe an advanced REVEALR platform, termed digital droplet REVEALR (ddREVEALR), that can achieve direct viral detection and absolute sample quantitation utilizing a signal amplification strategy that relies on chemical modifications, DNAzyme multiplexing, and volume compression. Using an AI-assisted image-based readout, ddREVEALR was found to achieve 95% positive predictive agreement from a set of 20 nasal pharyngeal swabs collected at UCI Medical Center in Orange, California. We propose that the combination of amplification-free and protein-free analysis makes ddREVEALR a promising application for direct viral RNA detection of clinical samples.
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