Abstract
Quantitative measurements of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in raw wastewater have been implemented worldwide since the beginning of the pandemic. Recent efforts are being made to evaluate different viral concentration methodologies to overcome supplier shortages during lockdowns. A set of 22-wastewater samples seeded with murine hepatitis virus (MHV), a member of the Coronaviridae family, and the bacteriophage MS2, were used to characterize and compare two ultrafiltration-based methods: a centrifugal ultrafiltration device (Centricon® Plus-70) and the automated concentrating pipette CP-Select™. Based on the recovery efficiencies, significant differences were observed for MHV, with Centricon® Plus-70 (24%) being the most efficient method. Nevertheless, concentrations of naturally occurring SARS-CoV-2, Human adenoviruses and JC polyomaviruses in these samples did not result in significant differences between methods suggesting that testing naturally occurring viruses may complement the evaluation of viral concentration methodologies. Based on the virus adsorption to solids and the necessity of a pre-centrifugation step to remove larger particles and avoid clogging when using ultrafiltration methods, we assessed the percentage of viruses not quantified after ultrafiltration. Around 23% of the detected SARS-CoV-2 would be discarded during the debris removal step. The CP-Select™ provided the highest concentration factor (up to 333×) and the lowest LoD (6.19 × 10 3 GC/l) for MHV and proved to be fast, automatic, highly reproducible and suitable to work under BSL-2 measures.
Keywords: SARS-CoV-2; Ultrafiltration; Viral concentration method; Viral recovery; Wastewater.
【저자키워드】 SARS-CoV-2, wastewater, Ultrafiltration, Viral concentration method, Viral recovery, 【초록키워드】 coronavirus, pandemic, Human, complement, severe acute respiratory syndrome Coronavirus, virus, Particle, wastewater, Measures, automated, methodology, MHV, murine hepatitis virus, Concentration, Ultrafiltration, Adsorption, lockdowns, acute respiratory syndrome, significant difference, acute respiratory syndrome coronavirus, significant differences, acute respiratory syndrome coronavirus 2, effort, Coronaviridae family, recent, BSL-2, lowest, highest, evaluate, provided, were used, overcome, the Coronaviridae, quantified, MS2, 【제목키워드】 Concentration,