A rapid, high-throughput, and sensitive PEG-precipitation method for SARS-CoV-2 wastewater surveillance

废水 离心 吞吐量 色谱法 环境科学 化学 计算机科学 环境工程 电信 无线
作者
Xiawan Zheng,Mengying Wang,Yu Deng,Xiaoqing Xu,Danxi Lin,Yulin Zhang,Shuxian Li,Jiahui Ding,Xianghui Shi,Chung In Yau,Leo L. M. Poon,Tong Zhang
出处
期刊:Water Research [Elsevier BV]
卷期号:230: 119560-119560 被引量:32
标识
DOI:10.1016/j.watres.2022.119560
摘要

The effective application of wastewater surveillance is dependent on testing capacity and sensitivity to obtain high spatial resolution testing results for a timely targeted public health response. To achieve this purpose, the development of rapid, high-throughput, and sensitive virus concentration methods is urgently needed. Various protocols have been developed and implemented in wastewater surveillance networks so far, however, most of them lack the ability to scale up testing capacity or cannot achieve sufficient sensitivity for detecting SARS-CoV-2 RNA at low prevalence. In the present study, using positive raw wastewater in Hong Kong, a PEG precipitation-based three-step centrifugation method was developed, including low-speed centrifugation for large particles removal and the recovery of viral nucleic acid, and medium-speed centrifugation for the concentration of viral nucleic acid. This method could process over 100 samples by two persons per day to reach the process limit of detection (PLoD) of 3286 copies/L wastewater. Additionally, it was found that the testing capacity could be further increased by decreasing incubation and centrifugation time without significantly influencing the method sensitivity. The entire procedure uses ubiquitous reagents and instruments found in most laboratories to obtain robust testing results. This high-throughput, cost-effective, and sensitive tool will promote the establishment of nearly real-time wastewater surveillance networks for valuable public health information.
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