Abstract
Wastewater-based epidemiology offers a cost-effective alternative to testing large populations for SARS-CoV-2 virus, and may potentially be used as an early warning system for SARS-CoV-2 pandemic spread. However, viruses are highly diluted in wastewater, and a validated method for their concentration and further processing, and suitable reference viruses, are the main needs to be established for reliable SARS-CoV-2 municipal wastewater detection. For this purpose, we collected wastewater from two European cities during the Covid-19 pandemic and evaluated the sensitivity of RT-qPCR detection of viral RNA after four concentration methods (two variants of ultrafiltration-based method and two adsorption and extraction-based methods). Further, we evaluated one external (bovine corona virus) and one internal (pepper mild mottle virus) reference virus. We found a consistently higher recovery of spiked virus using the modified ultrafiltration-based method. This method also had a significantly higher efficiency (p-value <0.01) for wastewater SARS-CoV-2 detection. The ultracentrifugation method was the only method that detected SARS-CoV-2 in the wastewater of both cities. The pepper mild mottle virus was found to function as a potentially suitable internal reference standard.
Keywords: Bovine corona virus; Municipal wastewater; Pepper mild mottle virus; RT-qPCR; SARS-CoV-2; Virus concentration method.
【저자키워드】 SARS-CoV-2, RT-qPCR, Pepper mild mottle virus, Bovine corona virus, Municipal wastewater, Virus concentration method., 【초록키워드】 viruses, pandemic, Epidemiology, SARS-CoV-2 pandemic, COVID-19 pandemic, variant, SARS-CoV-2 virus, virus, Population, Spread, SARS-CoV-2 detection, sensitivity, RT-qPCR, wastewater, Viral, qPCR, Mild, bovine, Viral RNA, reference standard, Concentration, Ultrafiltration, Adsorption, Efficiency, P-value, ultracentrifugation, offer, European, collected, evaluated, significantly higher, 【제목키워드】 virus, quantification, Concentration, Benchmarking,