Wastewater Concentration using the Electronegative Filtration method, Total Nucleic Acid Extraction and PCR inhibitor removal for recovery of viral pathogens v1

This standard operating procedure (SOP) sets out the workflow for processing wastewater samples with the goal of recovering and detecting viral pathogens. The steps include virus concentration using the electronegative filtration (ENF) method, total nucleic acid extraction using MagMAX Wastewater Ultra Nucleic Acid Isolation Kit, and PCR Inhibitor removal using the Zymo OneStep PCR Inhibitor Removal Kit. The raw wastewater/sewage samples are concentrated using the ENF method, which operates off the principle of adsorption-elution. The pH of the mixture is first adjusted, after which the sample is poured through a negatively charged membrane. Viruses are reversibly adsorbed to filters via charge attraction, after which they are eluted into a smaller volume. The resulting concentrate is then subjected to nucleic acid extraction and PCR inhibitor removal. All procedures can be performed by suitably trained members of staff.