Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA.
Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USA.
Nat Commun. 2021 May 18;12(1):2905. doi: 10.1038/s41467-021-23185-x.
The need for rapid, accurate, and scalable testing systems for COVID-19 diagnosis is clear and urgent. Here, we report a rapid Scalable and Portable Testing (SPOT) system consisting of a rapid, highly sensitive, and accurate assay and a battery-powered portable device for COVID-19 diagnosis. The SPOT assay comprises a one-pot reverse transcriptase-loop-mediated isothermal amplification (RT-LAMP) followed by PfAgo-based target sequence detection. It is capable of detecting the N gene and E gene in a multiplexed reaction with the limit of detection (LoD) of 0.44 copies/μL and 1.09 copies/μL, respectively, in SARS-CoV-2 virus-spiked saliva samples within 30 min. Moreover, the SPOT system is used to analyze 104 clinical saliva samples and identified 28/30 (93.3% sensitivity) SARS-CoV-2 positive samples (100% sensitivity if LoD is considered) and 73/74 (98.6% specificity) SARS-CoV-2 negative samples. This combination of speed, accuracy, sensitivity, and portability will enable high-volume, low-cost access to areas in need of urgent COVID-19 testing capabilities.
对于 COVID-19 诊断而言，快速、准确且可扩展的检测系统的需求是显而易见且紧迫的。在此，我们报告了一种快速可扩展便携检测（SPOT）系统，该系统由一种快速、高灵敏度且准确的检测方法和一个电池供电的便携式设备组成，可用于 COVID-19 的诊断。SPOT 检测方法包含了一步法逆转录环介导等温扩增（RT-LAMP），随后是基于 PfAgo 的靶序列检测。它能够在 30 分钟内对 SARS-CoV-2 病毒感染的唾液样本中的 N 基因和 E 基因进行多重反应检测，检测限（LoD）分别为 0.44 拷贝/μL 和 1.09 拷贝/μL。此外，该 SPOT 系统还用于分析 104 份临床唾液样本，确定了 28/30（93.3%的灵敏度）份 SARS-CoV-2 阳性样本（如果考虑到 LoD，则为 100%的灵敏度）和 73/74（98.6%的特异性）份 SARS-CoV-2 阴性样本。这种速度、准确性、灵敏度和便携性的结合将使人们能够以大容量、低成本的方式获得急需 COVID-19 检测能力的地区。
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