Meinig School of Biomedical Engineering, Cornell University, Ithaca, NY 14850, USA.
Department of Chemical, Environmental, and Materials Engineering, University of Miami, Coral Gables, FL 33146, USA.
Sci Total Environ. 2024 Sep 20;944:173744. doi: 10.1016/j.scitotenv.2024.173744. Epub 2024 Jun 4.
Quantification of SARS-CoV-2 RNA copies in wastewater can be used to estimate COVID-19 prevalence in communities. While such results are important for mitigating disease spread, SARS-CoV-2 measurements require sophisticated equipment and trained personnel, for which a centralized laboratory is necessary. This significantly impacts the time to result, defeating its purpose as an early warning detection tool. The objective of this study was to evaluate a field portable device (called MINI) for detecting SARS-CoV-2 viral loads in wastewater using real-time reverse transcriptase loop-mediated isothermal amplification (real-time RT-LAMP). The device was tested using wastewater samples collected from buildings (with 430 to 1430 inhabitants) that had known COVID-19-positive cases. Results show comparable performance of RT-LAMP against reverse transcriptase polymerase chain reaction (RT-qPCR) when detecting SARS-CoV-2 copies in wastewater. Both RT-LAMP and RT-qPCR detected SARS-CoV-2 in wastewater from buildings with at least three positive individuals within a 6-day time frame prior to diagnosis. The large 96-well throughput provided by MINI provided scalability to multi-building detection. The portability of the MINI device enabled decentralized on-site detection, significantly reducing the time to result. The overall findings support the use of RT-LAMP within the MINI configuration as an early detection system for COVID-19 infection using wastewater collected at the building scale.
污水中 SARS-CoV-2 RNA 拷贝的定量分析可用于估计社区内 COVID-19 的流行程度。虽然这些结果对于减轻疾病传播很重要,但 SARS-CoV-2 的测量需要复杂的设备和经过培训的人员,这需要一个集中的实验室。这会显著影响结果的时间,使其失去作为早期预警检测工具的意义。本研究的目的是评估一种现场便携式设备(称为 MINI),该设备使用实时逆转录环介导等温扩增(real-time RT-LAMP)检测污水中的 SARS-CoV-2 病毒载量。该设备使用从有已知 COVID-19 阳性病例的建筑物(有 430 至 1430 名居民)收集的废水样本进行了测试。结果表明,在检测污水中的 SARS-CoV-2 拷贝时,RT-LAMP 的性能与逆转录聚合酶链反应(RT-qPCR)相当。RT-LAMP 和 RT-qPCR 都能在诊断前 6 天内从至少有 3 名阳性个体的建筑物废水中检测到 SARS-CoV-2。MINI 提供的大型 96 孔通量为多建筑物检测提供了可扩展性。MINI 设备的便携性支持在 RT-LAMP 配置中使用,以使用建筑物规模收集的废水作为 COVID-19 感染的早期检测系统。