Defining the methodological approach for wastewater-based epidemiological studies—Surveillance​ of SARS-CoV-2

Abstract

Since COVID-19 outbreak, wastewater-based epidemiology (WBE) studies as surveillance system is becoming an emerging interest due to its functional advantage as a tool for early warning signal and to catalyze effective disease management strategies based on the community diagnosis. An attempt was made in this study to define and establish a methodological approach for conducting WBE studies in the framework of identifying/selection of surveillance sites, standardizing sampling policy, designing sampling protocols to improve sensitivity, adopting safety protocol, and interpreting the data. Data from hourly sampling indicated a peak in the viral RNA during the morning hours (6–9 am) when the all the domestic activities are maximum. The daily sampling and processing revealed the dynamic nature of infection spread among the population. The two sampling methods viz. grab, and composite showed a good correlation. Overall, this study establishes a structured protocol for performing WBE studies that could provide useful insights on the spread of the pandemic at a given point of time. Moreover, this framework could be extrapolated to monitor several other clinically relevant diseases. Following these guidelines, it is possible to achieve measurable and reliable SARS-CoV-2 RNA concentrations in wastewater infrastructure and therefore, provides a methodological basis for the establishment of a national surveillance system.

The Urban Water Cycle as a Planning Tool to Monitor SARS-CoV-2: A Review of the Literature

“…Countries such as the United States, Holland, Italy, Brazil, Spain, Australia and India have already benefitted from the implementation of surveillance protocols during this pandemic. Indeed, authors such as Kopperi et al [208] have already proposed a standard methodological approach for the study and epidemiological surveillance of SARS-CoV-2 in wastewater, that could be applied worldwide.…”

Longitudinal and Long-Term Wastewater Surveillance for COVID-19: Infection Dynamics and Zoning of Urban Community

“…An optimized sampling protocol was followed to sample the domestic wastewater at the selected points based on our earlier study [ 22 ]. The grab samples were collected in a clean plastic bottle (disposable; 1.2 L) containing 20 mL of sodium hypochlorite (0.1%) to inactivate the pathogens.…”

“…RNA was extracted from the concentrated samples using the Viral RNA isolation kit (Qiagen, Germantown, MD, USA) as per the provided manufacturer’s protocol. DNA/RNA cross-contamination was avoided by using sterile equipment and RNase-free water for the RNA extraction [ 22 , 28 ]. Isolated SARS-CoV-2 RNA was quantified by using the RT-PCR Detection Kit (Shanghai Fosun Long March Medical Science Co., Ltd., Shanghai, China) which is an FDA (Food and Drug Administration, USA Government) approved kit.…”

“…A load of viral material temporally alters sewage based on the time of defecation frequencies and sampling [ 19 , 20 ]. Diverse viral load shedding from the affected community, convergence of household wastewater and industrial effluents and time of sampling could affect the detection of the viral genome in the sewage; however, WBE provides a range of information to predict the dynamics of infection with the design of a sampling protocol [ 7 , 13 , 14 , 15 , 21 , 22 , 23 , 24 , 25 ]. The development of highly sensitive and specific RT-PCR tests to detect SARS-CoV-2 (the corona virus causing the COVID-19 pandemic) in sewage samples presents the potential to use wastewater sampling as a diagnostic test for SARS-CoV-2 in the community [ 21 , 22 , 23 , 24 ].…”

“…Different independent SARS-CoV-2 WBE studies have followed various sewage sampling and processing methods to detect the SARS-CoV-2 RNA in sewage [ 7 , 23 , 24 , 25 ]. The grab sampling method can be used in remote areas and even in poor sewage system conditions, which aids in improving surveillance [ 22 ]. These kinds of WBE methods will help to monitor surveillance in the area where proper hospitality infrastructure is lacking.…”