An optimized and robust PEG precipitation method for detection of SARS-CoV-2 in wastewater

Evaluation of SARS-CoV-2 concentrations in wastewater and river water samples

“…Therefore, using internal and external controls, specific methods should be validated at the time of the experiment for efficient recovery of SARS-CoV-2 from different types of wastewater and other water bodies. Based on our results and those of previous studies, PEG 8000 precipitation and ultrafiltration are efficient approaches for concentrating SARS-CoV-2 from wastewater and river water samples [ 16 , 17 , 20 ].…”

“…However, there are some limitations to WBE for SARS-CoV-2 surveillance, including the methodologies used to concentrate and recover the virus [ 15 ], the wastewater sampling source and volume (particles or aqueous portions) [ 6 , 16 , 17 ], the RNA extraction methods that lack a gold standard [ 17 ], the primers or molecular diagnosis strategy used [ 3 , 5 , 18 , 19 ], and the RT-qPCR kit used for testing [ 20 ]. In addition, the number of virus copies can vary depending on the recovery efficiencies of the methods [ 17 , 21 , 22 ].…”

High-Frequency, High-Throughput Quantification of SARS-CoV-2 RNA in Wastewater Settled Solids at Eight Publicly Owned Treatment Works in Northern California Shows Strong Association with COVID-19 Incidence

“…Due to the dilute nature of SARS-CoV-2 RNA in wastewater, most methods that focus on influent require a number of preanalytical steps to concentrate SARS-CoV-2 prior to extracting the viral RNA. Depending on the method, these steps include ultrafiltration ( 2 , 5 ), organic flocculation coupled to centrifugation ( 3 , 11 ), and charged membrane filtration ( 12 ). Although liquid wastewater monitoring can be utilized for large programs, each of these listed concentration steps add significant time, equipment, and personnel requirements to SARS-CoV-2 RNA quantification in wastewater.…”

High frequency, high throughput quantification of SARS-CoV-2 RNA in wastewater settled solids at eight publicly owned treatment works in Northern California shows strong association with COVID-19 incidence

“…Depending on the method, these steps include ultrafiltration 2,5 , organic flocculation coupled with centrifugation 3,11 , and charged membrane filtration 12 . Each of these listed concentration steps add significant time, equipment, and personnel requirements to SARS-CoV-2 RNA quantification in wastewater.…”