Comparison of high-frequency in-pipe SARS-CoV-2 wastewater-based surveillance to concurrent COVID-19 random clinical testing on a public U.S. university campus

Abstract

Wastewater-based epidemiology (WBE) is utilized globally as a tool for quantifying the amount of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) within communities, yet the efficacy of community-level wastewater monitoring has yet to be directly compared to random Coronavirus Disease of 2019 (COVID-19) clinical testing; the best-supported method of virus surveillance within a single population. This study evaluated the relationship between SARS-CoV-2 RNA in raw wastewater and random COVID-19 clinical testing on a large university campus in the Southwestern United States during the Fall 2020 semester. Daily composites of wastewater (24-hour samples) were collected three times per week at two campus locations from 16 August 2020 to 1 January 2021 ( n = 95) and analyzed by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) targeting the SARS-CoV-2 E gene. Campus populations were estimated using campus resident information and anonymized, unique user Wi-Fi connections. Resultant trends of SARS-CoV-2 RNA levels in wastewater were consistent with local and nationwide pandemic trends showing peaks in infections at the start of the Fall semester in mid-August 2020 and mid-to-late December 2020. A strong positive correlation ( r = 0.71 ( p < 0.01); n = 15) was identified between random COVID-19 clinical testing and WBE surveillance methods, suggesting that wastewater surveillance has a predictive power similar to that of random clinical testing. Additionally, a comparative cost analysis between wastewater and clinical methods conducted here show that WBE was more cost effective, providing data at 1.7% of the total cost of clinical testing ($6042 versus $338,000, respectively). We conclude that wastewater monitoring of SARS-CoV-2 performed in tandem with random clinical testing can strengthen campus health surveillance, and its economic advantages are maximized when performed routinely as a primary surveillance method, with random clinical testing reserved for an active outbreak situation.

Municipal and neighbourhood level wastewater surveillance and subtyping of an influenza virus outbreak

“…Since WWS is independent of clinical testing, it is not affected by such constraints and provides an unbiased representation of the temporal prevalence of the disease in this regard 40 . Hence, WWS of SARS-CoV-2 may provide similar predictive power to that of random clinical testing, and we hypothesize that the same holds true for influenza WWS 41 . Second, clinical test data is reported citywide by the date of clinical testing and not by the onset of symptoms, making it likely that fecal shedding of viral RNA could begin before individuals seek medical treatment and are clinically tested 42 , 43 .…”

Predicting COVID-19 Infected Individuals in a Defined Population from Wastewater RNA Data

“…For a specific community, an increase in viral RNA copies can then signal the need for more widespread clinical testing. To date, WBE has been employed in many venues ranging from small-scale communities, such as hospitals, nursing homes, airlines, and college campuses, to larger-scale populations, such as large towns and cities. − …”

Leveraging an established neighbourhood-level, open access wastewater monitoring network to address public health priorities: a population-based study

“…Samples were processed and analysed according to published studies. 5 , 14 , 15 Briefly, raw wastewater samples were analysed for SARS-CoV-2 following sequential steps of filtration (0·45 μm polyether sulfone membrane), viral concentration by ultracentrifugation (10 kDa molecular weight cutoff centrifugal filters), nucleic acid extraction (50 μL resultant RNA extract), and TaqMan-based (Invitrogen, Waltham, MA, USA) reverse transcription quantitative PCR (RT-qPCR) targeting the SARS-CoV-2 E (envelope) gene. Spike-and-recovery experiments were performed using murine hepatitis virus (as a surrogate) based on previously published methods.…”

“…Spike-and-recovery experiments were performed using murine hepatitis virus (as a surrogate) based on previously published methods. 5 , 16 , 17 Full method details for sample processing and analysis, including primer and probe sequences and RT-qPCR methods, are in the appendix (pp 2–3, 6–7 ).…”

“… 1 , 2 At the onset of the SARS-CoV-2 pandemic, substantial delays in conventional and individualised clinical testing, due in part to an overwhelmed health-care system and resource limitations, 3 positioned WBE as a promising supplemental tool for assessing SARS-CoV-2 spread at the population level. 4 , 5 , 6 , 7 Early data from the beginning of the pandemic suggested that SARS-CoV-2 levels in wastewater and sewage sludge offered early indication of clinical confirmed infections, disease, and mortality in a community. 8 , 9 …”