AI Biosciences, Inc. SBIR Phase I Award, September 2022

Project Summary/Abstract: 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.