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

使用高通量、高度多重数字阵列 PCR 技术提高废水中病原体检测（包括 SARS-CoV-2）的可扩展性、灵敏度和可解释性

• 批准号: 10320994
• 负责人: Rachel Todd Noble
• 金额: $ 67.33万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10320994
• 关键词: 2019-nCoV; Address; Biological Assay; Blood Circulation; Businesses; COVID-19; COVID-19 assay; COVID-19 pandemic; Clinical; Communicable Diseases; Communities; Consumption; Coronavirus; Data; Data Analyses; Development; Disease; Disease Surveillance; Education; Feces; Future; Geographic Locations; Geography; Goals; Health; Home; Hour; Human; Individual; Laboratories; Location; Methods; Molecular; Monitor; Pathogen detection; Patients; Performance; Plants; Play; Precipitation; Prevalence; Public Health; Quality Control; Reaction; Reporting; Research; Respiratory distress; Role; Sampling; Schools; Sewage; Source; System; Technology; Testing; Time; Vaccination; Vaccines; Virus; Vision; biomarker panel; community transmission; cost; digital; emerging pathogen; gastrointestinal; genetic testing; health management; health warning; improved; method development; molecular marker; multiplex assay; novel; novel coronavirus; pandemic disease; pathogen; research clinical testing; respiratory; response; tool; transmission process; viral RNA; wastewater epidemiology; wastewater monitoring; wastewater samples

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.

项目摘要 目前，RT-QPCR和数字PCR（DPCR）等分子技术的应用来量化 废水中的SARS-COV-2和相关目标繁琐，​​耗时且昂贵。尽管 在方法的开发和数据的解释方面取得了进展，还有很多事情仍然存在 可以改进该技术作为公共卫生管理工具。在 当前的方法是1）缺乏精简且一致的分析前处理步骤，2） 相关目标之间的覆盖范围需要从任何单个样本到 提供定量信息，3）缺乏关于病原体/标记面板的发展的视力， 就像在临床领域使用的那些一样，用于解释不同州，地区和地区的数据 国家。该项目的目的是成功地导航这三个限制 开发不依赖临床测试的公共卫生警告系统，具有能力 将来快速解决新颖的病原体威胁。

项目成果

Evaluating the impact of sample storage, handling, and technical ability on the decay and recovery of SARS-CoV-2 in wastewater.

• DOI: 10.1371/journal.pone.0270659
• 发表时间: 2022
• 期刊: PLOS ONE
• 影响因子: 3.7
• 作者: Beattie, Rachelle E.;Blackwood, A. Denene;Clerkin, Thomas;Dinga, Carly;Noble, Rachel T.
• 通讯作者: Noble, Rachel T.