Streamlining RNA extraction for widespread COVID-19 diagnostic testing

持续的COVID-19大流行凸显了广泛的诊断检测在追踪和遏制病毒传播方面的重要性。然而，在这次疫情期间，大规模检测的一个主要瓶颈是全球缺乏商业RNA提取试剂盒。这些试剂盒用于在检测前从患者样本中提取SARS-CoV-2病毒基因组，并且是大多数核酸检测测定的重要组成部分。RNA提取试剂盒的短缺严重推迟了全球几个地区的检测，并导致疫情爆发。加拿大尤其面临风险，因为它缺乏足够的国内RNA提取试剂盒制造商供应，其中大部分来自其他国家。依赖国际来源的解决方案面临更大的供应瓶颈风险，特别是在感染爆发期间。在这项提案中，Chou实验室与多伦多的分子诊断公司Luna Nanotech合作，开发一类新的核酸探针，简化RNA提取和检测，对现有解决方案进行三项改进：（i）不需要专用硬件的更简单和更快速的方案，（ii）通过靶特异性富集提高下游测定的检测灵敏度，以及（iii）更高的成本效率和可扩展性。我们将使用合成RNA、灭活病毒和患者样本开发和验证这项技术，为临床使用提供经过充分验证的试剂。该提案的结果将广泛支持核酸检测技术，从集中式RT-PCR检测、即时检测到下一代测序工作，通过提高检测能力和提高准确性。完成后，Luna Nanotech还将致力于将这种新的RNA提取技术与其现有的等温核酸检测分析管道相结合，用于即时检测。

The ongoing COVID-19 pandemic has highlighted the importance of widespread diagnostic testing in tracing and containing viral transmission. A major bottleneck to widespread testing during this outbreak, however, has been the global shortage of commercial RNA extraction kits. These kits are used to extract the SARS-CoV-2 viral genome from patient samples before testing, and are an essential component of the majority of nucleic-acid detection assays. Shortage of RNA extraction kits has critically delayed testing in several regions globally and led to outbreaks. Canada is especially at risk because it lacks sufficient domestic supply of RNA extraction kit manufacturers, and sources most of them from other countries. Reliance on internationally sourced solutions runs greater risk of a supply bottleneck, especially during periods of an infection outbreak. In this proposal, the Chou lab is partnering with Luna Nanotech, a Toronto-based molecular diagnostics company, to develop a new class of nucleic-acid probes that streamlines RNA extraction and detection, providing three improvements over existing solutions: (i) simpler and faster protocol that does not require specialty hardware, (ii) enhanced detection sensitivity for downstream assays via target-specific enrichment, and (iii) higher cost-efficiency and scalability. We will develop and validate this technology using synthetic RNA, deactivated viruses, and patient samples, providing a well-validated reagent for clinical use. The outcome of this proposal will broadly support nucleic-acid testing technologies from centralized RT-PCR testing, point-of-care assays, to next-generating sequencing efforts via improved testing capacity and enhanced accuracy. Following completion, Luna Nanotech will also aim to integrate this new RNA extraction technology with their existing pipeline of isothermal nucleic-acid detection assay for point-of-care testing.