Rapid Point-of-Care Molecular Test for SARS-CoV-2 and Influenza A/B

SARS-CoV-2 和甲型/乙型流感的快速护理点分子检测

• 批准号: 10484040
• 负责人: SEASON S-S WONG
• 金额: $ 27.58万
• 依托单位: AI BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10484040
• 关键词: Rapid; Point; Care; Molecular; Test

PROJECT SUMMARY The goal of this Phase I SBIR proposal is to develop a 15 minute multiplex point-of-care (POC) molecular test for the qualitative detection of COVID-19 and influenza A/B. As of September 2021, the US has had over 39.5 million confirmed cases of COVID-19 and over 642,000K deaths. Additionally, influenza viruses account for over 200,000 hospitalizations and 30,000–50,000 deaths in the US each year. As SARS-CoV-2 and influenza infections can hardly be differentiated by symptoms alone, a single test that can diagnose illness caused by SARS-CoV-2 and influenza viruses will give public health officials information they need in their efforts to control the spread of these important viruses of concern. Our approach leverages rapid sample preparation (1 min hands-on-time and 2.5 minute of heat inactivation) and the rapid gold-standard real-time RT-PCR (12 minute, 40-cycle) to amplify the targeted viral genome. This test can be utilized in multiple settings where actionable test results are needed to make informed treatment decisions quickly. We will demonstrate that our test can deliver rapid results with a similar level of performance as those obtained in reference labs using nasopharyngeal swabs and contrived saliva samples. Our approach has been tested using SARS-CoV-2 and influenza positive clinical specimens in viral transport media. We plan to achieve rapid multiplex RT-PCR using shuttle PCR with thin-film reactors. We can enhance PCR speed and efficiency by using vibration to enhance reagent mass-transport while avoiding the “extreme PCR” approach that needs 10 to 20 times more concentrated reagents (i.e., additional expense) to speed up the reaction. The result from spatial multiplexed fluorescence probe-based RT-PCR can be imaged and analyzed by on-board electronics. The RT-PCR test takes ~8 minutes to complete 40 cycles after 2.5 min of sample heat inactivation and 3 min of reverse-transcription step, and some high positive results can be obtained in as early as 6 minutes. Our approach will be much more sensitive than antigen-based approaches and faster than isothermal amplification approaches, such as loop mediated isothermal amplification (LAMP). The diagnostic utility of the molecular assay will be demonstrated by comparing its robustness, speed, sensitivity, and specificity with current molecular assays. In Phase I, we will work with our collaborator to validate our device and test using clinical specimens. If successfully developed, this compact and minimal instrumentation approach will be simple to perform and inexpensive enough for all sizes of primary-care physician’s offices, nursing homes, pharmacies, community health clinics, and even at-home consumers to adopt the platform.

项目概要 第一阶段 SBIR 提案的目标是开发 15 分钟多重即时护理 (POC) 分子测试 截至 2021 年 9 月，美国已检测出超过 39.5 例 COVID-19 和 A/B 型流感。 百万确诊的 COVID-19 病例和超过 642,000,000 人死亡。 由于 SARS-CoV-2 和流感，美国每年有 200,000 人住院治疗，30,000-50,000 人死亡。 仅凭症状很难区分感染，单一测试即可诊断由感染引起的疾病 SARS-CoV-2 和流感病毒将为公共卫生官员提供控制工作所需的信息 我们的方法利用快速样品制备（1 分钟）。 手动操作时间和 2.5 分钟热灭活）和快速黄金标准实时 RT-PCR（12 分钟， 40 个循环）以扩增目标病毒基因组。该测试可用于可操作测试的多种环境。 需要结果来快速做出明智的治疗决定，我们将证明我们的测试可以发挥作用。 快速获得结果，其性能水平与参考实验室使用鼻咽拭子获得的结果相似 我们的方法已经使用 SARS-CoV-2 和流感阳性进行了临床测试。 我们计划使用薄膜穿梭 PCR 来实现快速多重 RT-PCR。 我们可以通过使用振动来增强试剂传质，从而提高 PCR 速度和效率。 避免需要 10 至 20 倍浓缩试剂的“极端 PCR”方法（即额外的 费用）以加速反应，基于空间多重荧光探针的 RT-PCR 可以得到结果。 RT-PCR 测试需要大约 8 分钟才能完成 40 个循环。 2.5 分钟的样品热灭活和 3 分钟的反转录步骤，一些高阳性结果可以 我们的方法将比基于抗原的方法灵敏得多。 并且比环介导等温扩增 (LAMP) 等等温扩增方法更快。 分子检测的诊断效用将通过比较其稳健性、速度、灵敏度、 在第一阶段，我们将与我们的合作者合作来验证我们的设备。 如果开发成功，这种紧凑且最小的仪器方法将使用临床标本进行测试。 操作简单且价格低廉，适合各种规模的初级保健医生办公室、疗养院、 药店、社区卫生诊所、甚至家庭消费者都采用该平台。