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Room Temperature Isothermal Colorimetric Padlock Probe Rolling Circle Amplification for Viral RNA Detection
bioRxiv - Synthetic Biology Pub Date : 2021-07-17 , DOI: 10.1101/2020.06.12.128876
Wilson Huang , Hannah Hsu , Jimmy Su , Jude Clapper , Jonathan Hsu

Seasonal flu and pandemics, which account for millions of infections and hundreds of thousands of deaths, require rapid and reliable detection mechanisms for preventive and therapeutic measures. Current methods of viral detection have limitations in speed, accuracy, accessibility, and usability. This project presents a novel, widely applicable viral diagnosis that uses a modified version of the traditional rolling circle amplification (RCA) to be sensitive, specific, direct, colorimetric, and operable at room temperature. We are specifically aiming to detect SARS-CoV-2, Influenza A (H1N1pdm09), and Influenza B (Victoria Lineage). Results using synthetic viral DNA sequences show that the diagnostic test could take as fast as 30 minutes and detect up to picomolar concentrations of DNA strands. The next step for this project is to test the assay with synthetic viral RNA to verify the results. We envision that the implementation of this type of diagnostic test could allow faster responses to outbreaks of related viruses and quicker societal recovery.

中文翻译:

用于病毒 RNA 检测的室温等温比色挂锁探针滚环扩增

季节性流感和大流行导致数百万感染和数十万人死亡,因此需要快速可靠的检测机制来采取预防和治疗措施。当前的病毒检测方法在速度、准确性、可访问性和可用性方面存在局限性。该项目提出了一种新的、广泛适用的病毒诊断,它使用传统滚环扩增 (RCA) 的改进版本,具有灵敏、特异、直接、比色和可在室温下操作的特点。我们的目标是检测 SARS-CoV-2、甲型流感 (H1N1pdm09) 和乙型流感 (Victoria Lineage)。使用合成病毒 DNA 序列的结果表明,诊断测试可能需要 30 分钟,并检测出高达皮摩尔浓度的 DNA 链。该项目的下一步是用合成病毒 RNA 测试该测定以验证结果。我们设想实施这种类型的诊断测试可以更快地应对相关病毒的爆发和更快的社会恢复。
更新日期:2021-07-19
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