Streamlining RNA extraction for widespread COVID-19 diagnostic testing

正在进行的新冠肺炎大流行突显了广泛的诊断检测在追踪和控制病毒传播方面的重要性。然而，这次暴发期间广泛检测的一个主要瓶颈是全球商业RNA提取试剂盒的短缺。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，是大多数核酸检测分析的重要组成部分。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。加拿大面临的风险尤其大，因为它国内缺乏足够的RNA提取试剂盒制造商，而且大部分都是从其他国家采购的。依赖国际来源的解决方案有更大的供应瓶颈风险，特别是在感染暴发期间。在这项提议中，周实验室将与多伦多的分子诊断公司露娜纳米技术公司合作开发一种新型核酸探针，以简化RNA提取和检测，在现有解决方案的基础上提供三项改进：(I)不需要特殊硬件的更简单、更快的协议，(Ii)通过靶标特定浓缩提高下游分析的检测灵敏度，以及(Iii)更高的成本效益和可扩展性。我们将使用合成RNA、灭活病毒和患者样本来开发和验证这项技术，为临床使用提供一种经过充分验证的试剂。这项提案的结果将广泛支持核酸测试技术，从集中式RT-PCR测试、护理点分析，到通过改进测试能力和提高准确性的下一代测序工作。完成后，露娜纳米科技还将致力于将这项新的核糖核酸提取技术与他们现有的用于护理点检测的等温核酸检测流水线相结合。

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.