Wastewater Assessment for Coronavirus in Kentucky: Implementing Enhanced Surveillance Technology

肯塔基州冠状病毒废水评估--实施强化监测 技术 对SARS-CoV-2的监测受到可用检测的阻碍，特别是在偏远和农村 区域。废水中SARS-CoV-2病毒生物标志物的筛选为个体提供了一种可行的替代方案 测试，它可以识别有可能成为热点的社区和设施。废水 监测克服了临床监测的几个局限性，例如需要强大的医疗保健 和实验室基础设施，以及缺乏代表性和全面的测试 社区。常规废水监测从下水道系统或废水中采集样本 处理设施，并在先进的PCR技术之前使用一系列提取步骤来定量 病毒生物标记物(RNA)。这种方法耗费时间和资源，这限制了它的广泛应用 申请。开发下一代技术以简化废水RNA提取和 量化将使其在设施和农村社区更广泛地使用是可行的。有限的 肯塔基州东南部农村地区新冠肺炎的临床检测阻碍了疾病监测和预防 在知情的情况下采取公共行动，减轻和控制疾病的传播。SARS冠状病毒废水检测 在这些社区中，使用现场友好型技术将为当地提供重要信息 当局和市民了解SARS-CoV-2感染在其社区的传播和趋势。我们的 该项目将实现两个目标：1)开发下一代废水评估技术；2) 实施和评估下一代废水检测。对于目标1，我们采用了发明的技术 由我们的团队称为基于排除的样本准备(ESP)，以简化和改进RNA提取 从废水中排出。我们将ESP与环介导的等温放大(LAMP)技术配对，以实现 RNA检测创建了一个敏感、强大和现场友好的平台，用于测试SARS的废水- CoV-2核糖核酸。我们将比较下一代化验与现有技术在衡量 灵敏度、特异度和可用性(例如，分析时间、分析步骤数)。对于目标2，我们将首先 在聚集的生活设施中以并排比较的方式验证下一代测试 与传统的废水监测相结合。接下来，在阿巴拉契亚地区建立现有的关系 肯塔基州，我们将招募和培训一个有目的的污水处理厂操作员小组，分水岭 观看公民科学家和学校科学教师在他们的社区和学校测试废水 使用野外友好的下一代废水检测。现场结果将在实验室中进行验证。一个 使用RE-AIM框架的稳健混合方法评估将评估社区对 SARS-CoV-2污水监测和社区识别的可行性、可接受性和实用性 针对检测结果采取的措施。

Wastewater Assessment for Coronavirus in Kentucky – Implementing Enhanced Surveillance Technology Surveillance for SARS-CoV-2 is hindered by the availability of testing, particularly in remote and rural areas. Screening of wastewater for SARS-CoV-2 viral biomarkers offers a viable alternative to individual testing and it can identify communities and facilities that are at risk of becoming hotspots.Wastewater surveillance overcomes several limitations of clinical surveillance, such as the need for robust healthcare and laboratory infrastructure and the lack of representative and comprehensive testing within communities. Conventional wastewater surveillance takes samples from sewer systems or wastewater treatment facilities and uses a series of extraction steps prior to advanced PCR technology to quantitate the viral biomarker (RNA). This approach is time and resource-intensive, which limits its wide-scale application. Developing next generation technology to simplify wastewater RNA extraction and quantitation will make it feasible to use more broadly at facilities and in rural communities. The limited clinical testing for COVID-19 in rural Southeastern Kentucky hampers disease surveillance and prevents informed public action to mitigate and contain the spread of disease. Wastewater testing for SARS-CoV- 2 in these communities using field-friendly technology will provide important information to local authorities and citizens about the spread and trend of SARS-CoV-2 infection in their communities. Our project will accomplish two aims: 1) Develop next generation wastewater assessment technology and 2) Implement and evaluate the next generation wastewater assay. For Aim 1 we adapt technology invented by our team termed exclusion-based sample preparation (ESP) to simplify and improve RNA extraction from wastewater. We will pair ESP with loop-mediated isothermal amplification (LAMP) technology for RNA detection to create a sensitive, robust, and field-friendly platform for testing wastewater for SARS- CoV-2 RNA. We will compare the next generation assay with established techniques on metrics of sensitivity, specificity, and usability (e.g., assay time, number of assay steps). For Aim 2 we will first validate the next generation assay in the field at congregate living facilities in a side-by-side comparison with conventional wastewater surveillance. Next, building on existing relationships in Appalachian Kentucky, we will recruit and train a purposive group of wastewater treatment plant operators, watershed watch citizen scientists, and school science teachers to test wastewater in their communities and schools using the field-friendly next generation wastewater assay. Field results will be validated in the lab. A robust mixed methods evaluation using the RE-AIM framework will assess community perceptions of feasibility, acceptability, and utility of wastewater surveillance for SARS-CoV-2 and identify community measures taken in response to test results.