Evaluation of Pre-Analytical and Analytical Methods for Detecting SARS-CoV-2 in Municipal Wastewater Samples in Northern Italy

(1) Background: The surveillance of SARS-CoV-2 RNA in urban wastewaters allows one to monitor the presence virus a population, including asymptomatic and symptomatic individuals, capturing real circulation this pathogen. aim study was evaluate performance different pre-analytical analytical methods for identifying untreated municipal samples by conducting an inter-laboratory proficiency test. (2) Methods: three concentration, namely, (A) Dextran PEG-6000 two-phase separation, (B) PEG-8000 precipitation without chloroform purification step (C) with were combined protocols extraction using commercial kits tested two primers/probe sets master mixes. (3) Results: treatment showed best recovery; no major differences observed among protocol one-step real-time RT-PCR mix kits. highest analytic sensitivity targeting N1/N3 fragments SARS-CoV-2. (4) Conclusions: combination N gene (two fragments) performing workflow detection wastewaters.