Comparison of Nanotrap® Microbiome A Particles, membrane filtration, and skim milk workflows for SARS-CoV-2 concentration in wastewater

Introduction Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) RNA monitoring in wastewater has become an important tool for Coronavirus Disease 2019 (COVID-19) surveillance. Grab (quantitative) and passive samples (qualitative) are two distinct sampling methods. Although many viral concentration methods such as the usage of membrane filtration skim milk reported, these generally require large volumes wastewater, expensive lab equipment, laborious processes. Methods The objectives this study were to compare workflows (Nanotrap ® Microbiome A Particles coupled with MagMax kit RNeasy kit) SARS-CoV-2 recovery grab Moore swab samples. Nanotrap particle workflow was initially evaluated without addition enhancement reagent 1 (ER1) 10 mL wastewater. RT-qPCR targeting nucleocapsid protein used detecting RNA. Results Adding ER1 prior significantly improved results ( P < 0.0001) processed by automated or manual workflows. concentrations a whole showed higher 0.001) than 150 using 250 workflow, respectively. Spiking known genome copies (GC) inactivated into indicated that limit detection ~11.5 GC/mL 115 digital PCR Discussion These suggest could substitute traditional compromising assay sensitivity. can be resource-limited areas, is appropriate large-scale COVID-19 wastewater-based