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.

项目摘要 该I期SBIR提案的目标是开发一种15分钟的多重床旁（POC）分子检测 用于COVID-19和甲型/B流感的定性检测。截至2021年9月，美国已有超过39.5 2019冠状病毒病确诊病例达100万例，死亡人数超过64.2万。此外，流感病毒占 美国每年有20万人住院，3万至5万人死亡。作为SARS-CoV-2和流感 感染很难单独通过症状来区分，一个单一的测试可以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他们在努力控制 这些重要病毒的传播。我们的方法利用快速样品制备（1分钟 手工操作时间和2.5分钟热灭活）和快速金标准实时RT-PCR（12分钟， 40个循环）以扩增靶向病毒基因组。该测试可用于多种设置，其中可操作的测试 需要有结果才能迅速作出知情的治疗决定。我们将证明我们的测试可以提供 快速结果，性能水平与参考实验室使用鼻咽拭子获得的结果相似 和人工唾液样本我们的方法已经使用SARS-CoV-2和流感阳性临床试验进行了测试。 病毒运输培养基中的标本。我们计划利用穿梭PCR和薄膜PCR实现快速多重RT-PCR， 反应堆我们可以通过使用振动来增强试剂的质量传输， 避免了需要10到20倍浓缩试剂的“极端PCR”方法（即，额外 （二）加快反应速度。基于空间多重荧光探针的RT-PCR的结果可以 通过机载电子设备进行成像和分析。RT-PCR检测需要约8分钟完成40个循环， 2.5样品热灭活3 min和逆转录步骤3 min，一些高阳性结果可 最早可在6分钟内获得。我们的方法将比基于抗原的方法灵敏得多 并且比等温扩增方法，如环介导等温扩增（LAMP）更快。 将通过比较其稳健性、速度、灵敏度， 和特异性。在第一阶段，我们将与我们的合作者一起验证我们的设备。 并使用临床样本进行测试。如果开发成功，这种紧凑和最小化的仪器方法 操作简单，价格低廉，适用于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，甚至在家里的消费者采用该平台。