SBIR Phase I: Automated In-Situ High-Resolution COVID-19 Wastewater-Based Epidemiology

SBIR第一阶段项目的更广泛影响是，通过基于废水的流行病学对现有和新出现的对公共卫生构成高风险的病毒进行监测。随着大流行形势的迅速演变以及最近冠状病毒变体的出现，人们普遍担心病毒变体比其前身更具传染性，并且可能在没有早期发现和预警的情况下在社区内迅速传播。其他令人关切的问题是，现有疫苗可能无法覆盖未来的突变，在这种情况下，通过基于废水的高分辨率流行病学进行早期监测将成为对当地社区内存在这种突变病毒变体提供早期警报的唯一有效手段之一。随着市政当局、大学校园和老年护理机构等大小社区广泛采用拟议的仪器，该项目能够在大流行期间早期发现和监测当前的冠状病毒及其变种。该技术还可以推广到今后几年有可能在社区传播的其他新出现的病毒，不仅适用于废水，还适用于广泛的环境和娱乐水质监测。拟建项目是一种高度优化的逆转录定量聚合酶链反应（RT-qPCR）方法，用于原位采样和分析，并实现相应的仪器平台，适用于不同病毒的早期检测和监测，包括当前冠状病毒（SARS-CoV-2）及其变体，以及其他新兴病毒。与目前基于废水的流行病学方法相比，该技术旨在缩短获得结果的时间，同时最大限度地降低成本和物流。这项工作将包括测试几种可修改的方案，以适应自动原位SARS-CoV-2测量，然后选择和优化最合适的方案。随后将使用加标样品以及适当的阳性和阴性对照对方案进行测试和验证。该方法将首次在自动化设备中实现分子生物学协议的最新进展，例如无提取的单步RT-qPCR。该项目致力于实现第一个可用的原位RT-qPCR，有可能彻底改变基于废水的流行病学，一般环境测试和来源跟踪领域。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact of this SBIR Phase I project addresses the monitoring needs via wastewater-based epidemiology of current and emerging viruses that pose high risks to public health. With the rapidly evolving nature of the pandemic situation and the recent emergence of the coronavirus variants, one common concern is that the virus variants are more contagious than their predecessors, and will likely spread quickly within communities without early detection and warning. Other concerns are that future mutations may not be covered by existing vaccines, in which case early monitoring through high resolution wastewater-based epidemiology will become one of the only effective means of providing early alerts of the presence of such mutated virus variants within local communities. With a wide adaptation of the proposed instrumentation by municipalities, large and small communities such as college campuses and senior care facilities, the project enables the early detection and monitoring of the current coronavirus and its variants during the pandemic. The technology can also be extended to other emerging viruses with risks of community spread in the years to follow, not only in wastewater but also in widespread environmental and recreational water quality monitoring.The proposed project is a highly-optimized reverse transcription quantitative polymerase chain reaction (RT-qPCR) approach for in-situ sampling and analysis, and the realization of a corresponding instrumentation platform that is applicable to the early detection and monitoring of different viruses, including the current coronavirus (SARS-CoV-2), its variants, and other emerging viruses. The proposed technology aims to reduce the time-to-result compared to current wastewater-based epidemiology approaches while minimizing cost and logistics. The effort will include the testing of several protocols that are amendable to adaption for automated in-situ SARS-CoV-2 measurement, followed by the selection and optimization of the most suitable one. The protocols will be subsequently tested and validated using spiked samples, along with appropriate positive and negative controls. The approach will implement, for the first time in an automated device, the latest advances in molecular biology protocols, such as extraction-free single-step RT-qPCR. The project strives to achieve the first in-situ RT-qPCR available, with the potential to completely revolutionize the fields of wastewater-based epidemiology, general environmental testing and source tracking.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.