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，实时定量聚合酶链式反应(RT-qPCR)和数字聚合酶链式反应(DPCR)等分子技术的应用 废水中的SARS-CoV-2和相关目标是繁琐、耗时和昂贵的。而当 在方法的发展和数据的解释方面已经取得了进展，还有很多事情要做 改进这项技术，将其用作公共卫生管理工具。它的一个主要缺点是 目前的方法是：1)缺乏简化和一致的分析前处理步骤；2) 覆盖相关目标需要从任何单个样本到 提供定量信息，以及3)对病原体/标记物小组的发展缺乏远见， 非常类似于临床领域中使用的，用于解释不同州、地区和 国家。本项目的目标将是成功地驾驭这三个限制 开发不依赖临床检测的公共卫生预警系统，并具有 以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.