Optimization and comparative study of SARS-CoV-2 RNA extraction principles for enhanced viral RNA yield in wastewater...

Detecting SARS-CoV-2 in wastewater provides a cost-effective alternative to expensive methods like random group testing and individual clinical tests. The method also allows public health authorities to account for asymptomatic cases because wastewater surveillance does not require individual opt-in to succeed, nor does it generate personally identifying information requiring a complex institutional review board (IRB) protocol when used at a community scale. An electronegative membrane filter (EMF) is a widely used, cost-effective tool for large-volume virus concentration. Here, we evaluate the efficiency of several widely available RNA extraction kits for wastewater surveillance, when used in combination with the EMF method of virus concentration. Two RNA extraction kits using lysis buffer protocols and two using bead-beating protocols were optimized and compared. The Zymo Quick RNA Viral Kit outperformed the other three kits in all performance metrics used to evaluate and was also the most cost-effective of the kits evaluated. In this study, we report detailed results of our independent evaluation of these kits and resources to aid researchers and public health agencies in their optimization of wastewater surveillance workflows.