This study presents the results of systematic wastewater monitoring of SARS-CoV-2 RNA and basic wastewater parameters from four different wastewater treatment plants (WWTPs) in the Czech Republic over the 2020-2022 epidemic. Two-step reverse-transcription quantitative PCR targeting genes encoding the N and Nsp12 proteins was employed to detect SARS-CoV-2 RNA loading in 420 wastewater samples. The results obtained were used to evaluate the potential of wastewater analysis for describing the epidemiological situation in cities of different sizes and determining temporal differences based on the prevailing SARS-CoV-2 variant. Strong correlations between the number of active and hospitalised COVID-19 cases in each WWTP catchment area and the concentration of SARS-CoV-2 RNA detected in the wastewater clearly demonstrated the suitability of this wastewater-based epidemiological approach for WWTPs of different sizes and characteristics, despite differences in SARS-CoV-2 variant waves, with some WWTPs showing high predictive potential. This study demonstrated on the data from the Czech Republic that targeted systematic monitoring of wastewater provides sufficiently robust data for surveillance of viral loads in sample populations, and thus contributes to preventing the spread of infection and subsequent introduction of appropriate measures.

• Keywords
• COVID-19, PCR, Virus, Wastewater treatment plant, Wastewater-based epidemiology,
• Publication type
• Journal Article MeSH

Many reports have documented that the presence of SARS-CoV-2 RNA in the influents of municipal wastewater treatment plants (WWTP) correlates with the actual epidemic situation in a given city. However, few data have been reported thus far on measurements upstream of WWTPs, i.e. throughout the sewer network. In this study, the monitoring of the presence of SARS-CoV-2 RNA in Prague wastewater was carried out at selected locations of the Prague sewer network from August 2020 through May 2021. Various locations such as residential areas of various sizes, hospitals, city center areas, student dormitories, transportation hubs (airport, bus terminal), and commercial areas were monitored together with four of the main Prague sewers. The presence of SARS-CoV-2 RNA was determined by reverse transcription - multiplex quantitative polymerase chain reaction (RT-mqPCR) after the precipitation of nucleic acids with PEG 8,000 and RNA isolation with TRIzol™ Reagent. The number of copies of the gene encoding SARS-CoV-2 nucleocapsid (N1) per liter of wastewater was compared with the number of officially registered COVID-19 cases in Prague. Although the data obtained by sampling wastewater from the major Prague sewers were more consistent than those obtained from the small sewers, the correlation between wastewater-based and clinical-testing data was also good for the residential areas with more than 7,000 registered inhabitants. It was shown that monitoring SARS-CoV-2 RNA in wastewater sampled from small sewers could identify isolated occurrences of COVID-19-positive cases in local neighborhoods. This can be very valuable while tracking COVID-19 hotspots within large cities.

• Keywords
• COVID-19 epidemic, Prague, RT-mqPCR, SARS-CoV-2, Wastewater-based epidemiology,
• MeSH
• Water Purification * MeSH
• COVID-19 * epidemiology   MeSH
• Humans MeSH
• Wastewater MeSH
• RNA, Viral MeSH
• SARS-CoV-2 MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Waste Water MeSH
• RNA, Viral MeSH

The COVID-19 pandemic has revealed a significant number of cracks in the current vigilance techniques that stand to minimise outbreaks of SARS-CoV-2. There is a serious inadequacy of the testing capacity of healthcare systems worldwide, which can be attributed to the lack of appropriate testing and monitoring methods for a disease such as COVID-19. The current tools in use for COVID-19 surveillance are either expensive, not applicable to large populations or yield results after the outbreak has already occurred. The immense contagiousness in combination with a wealth of asymptomatic carriers means that RT-PCR testing is not feasible on a mass scale. It is evident that new methods are required for the monitoring of COVID-19 and a range of new epidemiological tools must be implemented if public health systems worldwide want to make relevant predictions on the patterns of disease spread and increase the efficacy of their decisions. In addition to this, the pandemic has highlighted the necessity for redirecting biomedical research towards early diagnosis and rational therapy of respiratory viruses in particular, as well as prevention of their spread by conventional means. An efficient early detection system would save lives and allow countries to return to pre-pandemic standards of living. At the forefront of this lies wastewater-based epidemiology, which carries immense potential as a means of pre-symptomatic diagnosis and population-based surveillance.

• Keywords
• COVID-19, PCR, SARS-CoV-2, epidemiology, outbreaks, prevention, surveillance, wastewater-based epidemiology,
• MeSH
• COVID-19 * epidemiology   MeSH
• Disease Outbreaks prevention & control   MeSH
• Wastewater-Based Epidemiological Monitoring MeSH
• Humans MeSH
• Wastewater MeSH
• Pandemics prevention & control   MeSH
• SARS-CoV-2 MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Waste Water MeSH

New methodologies based on the principle of "sewage epidemiology" have been successfully applied before in the detection of illegal drugs. The study describes the idea of early detection of a virus, e.g., SARS-CoV-2, in wastewater in order to focus on the area of virus occurrence and supplement the results obtained from clinical examination. By monitoring temporal variation in viral loads in wastewater in combination with other analysis, a virus outbreak can be detected and its spread can be suppressed early. The use of biosensors for virus detection also seems to be an interesting application. Biosensors are highly sensitive, selective, and portable and offer a way for fast analysis. This manuscript provides an overview of the current situation in the area of wastewater analysis, including genetic sequencing regarding viral detection and the technological solution of an early warning system for wastewater monitoring based on biosensors.

• Keywords
• COVID-19, PCR methods, SARS-CoV-2, biosensors, genetic sequencing, virus detection, wastewater monitoring,
• MeSH
• COVID-19 * MeSH
• Wastewater-Based Epidemiological Monitoring * MeSH
• Humans MeSH
• Mutation MeSH
• Wastewater MeSH
• Sewage MeSH
• SARS-CoV-2 MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Names of Substances
• Waste Water MeSH
• Sewage MeSH