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SARS-CoV-2 is a pathogenic strain of coronavirus which causes acute respiratory disease in humans. Community spread of COVID-19 was difficult to assess in the beginning of the pandemic, so new methods of detecting the virus had to be discovered. For this reason, national wastewater surveillance systems were implemented in order to detect the spread of SARS-CoV-2 virus. However, to establish such systems, there’s a need to standardize protocols for coronavirus concentration, as well as finding the optimal titre of BCoV, generally used as a recovery control. In our study, we used a strain of Betacoronavirus 1 (EVAg 015V-02282) provided by EVAg as our process control. In order to set up which concentration of the Bovine coronavirus (BCoV) will give the most conclusive results, we diluted the virus three times by the decimal method before running the samples in the QIAcuity Digital PCR (dPCR). The evaluation was made so that the BCoV concentration could be used for future studies.

Wastewater-based epidemiology (WBE) has been previously used as a tool for pathogen identification within communities. After the SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) outbreak, in 2020, Daughton proposed the implementation of a wastewater surveillance strategy that could determine the incidence of COVID-19 (coronavirus disease 2019) nationally. Individuals in various stages of SARS-CoV-2 infection, including presymptomatic, asymptomatic and symptomatic patients, can be identified as carriers of the virus in their urine, saliva, stool and other bodily secretions. Studies using this method were conducted to monitor the prevalence of the virus in high-density populations, such as cities but also in smaller communities, such as schools and college campuses. The aim of this pilot study was to assess the feasibility and effectiveness of wastewater surveillance in Bucharest, Romania, and wastewater samples were collected weekly from seven locations between July and September 2023. RNA (ribonucleic acid) extraction, followed by dPCR (digital polymerase chain reaction) analysis, was performed to detect viral genetic material. Additionally, NGS (next generation sequencing) technology was used to identify the circulating variants within the wastewater of Bucharest, Romania. Preliminary results indicate the successful detection of SARS-CoV-2 RNA in wastewater, providing valuable insights into the circulation of the virus within the community., (© 2024. The Author(s).)