Improved scalability, sensitivity, and interpretability of pathogen detection, including SARS-CoV-2, in wastewater using high-throughput, highly multiplexed digital array PCR technology

项目摘要 目前，应用分子技术如RT-qPCR和数字PCR（dPCR）来定量 废水中的SARS-CoV-2和相关靶标是繁琐、耗时且昂贵的。而 在方法的发展和数据的解释方面已经取得了进展，但仍有许多工作要做。 该技术将作为公共卫生管理工具加以改进。一个主要的缺点， 目前的方法是1）缺乏精简和一致的分析前处理步骤，2） 覆盖相关靶标需要来自任何单个样品的大量反应（>20）， 提供定量信息，和3）缺乏对病原体/标志物小组发展的远见， 就像临床领域使用的那些，用于解释不同州、地区和 nations.本项目的目标是成功地驾驭这三个限制， 开发一个公共卫生预警系统，该系统不依赖于临床测试， 来快速应对未来新的病原体威胁。

PROJECT ABSTRACT Presently, the application of molecular technology such as RT-qPCR and digital PCR (dPCR) to quantify SARS-CoV-2 and related targets in wastewater is cumbersome, time consuming, and costly. While progress has been made on the development of methods and the interpretation of data, much remains to be improved for the technology to be used as a public health management tool. A major drawback in the current approaches are 1) the lack of streamlined and consistent pre-analytical processing steps, 2) coverage across the relevant targets requires a high number of reactions (>20) from any single sample to provide quantitative information, and 3) a lack of vision on the development of a pathogen/marker panel, much like those used in clinical arenas, for interpretation of the data across different states, regions and nations. The goal of this project will be to successfully navigate these three limitations toward development of a public health warning system that is not dependent on clinical testing and has the ability to rapidly address novel pathogen threats in the future.