Eurofins Covid-19 Sentinel™ Wastewater Test Provide Early Warning of a potential COVID-19 outbreak

Abstract

<jats:p>The Eurofins Covid-19 SentinelTM program was developed to monitor the evolution of the pandemic and for early detection of outbreaks. The study objective was to develop a wastewater testing method to analyze SARS-CoV-2 as an indicator of community infection rate resurgence of COVID-19 or in well-defined sites such as production facilities or nursing homes. Eurofins performed &gt;700 tests on 78 unique samples from 18 sites in Denmark, France and Belgium. Ten variant test protocols were trialed. Protocol variations trialed included centrifugation, precipitation of the SARS-CoV-2 RNA, agitation prior to precipitation, cooling, and pasteurization of the samples. A method was succesfully developed and reliability was supported by stability, reproducibility, and dilution &amp; linearity studies. Results obtained showed a direct link to number of RNA copies in the sample using a calibration curve with synthetic SARS-CoV-2. Analysis was performed on both the liquid phase and solid phase of wastewater samples, with virus RNA detected in both phases but more frequently in the liquid phase. The virus was present in a sample from a Danish community wastewater treatment plant collected on February 24, 3 days before the first COVID-19 case was officially reported in the country. The greatest concentration of virus detected corresponded to when the COVID-19 crisis was at its peak in Denmark. Based on studies carried out in a Danish hospital, the wastewater testing method is expected to be able to detect a community COVID-19 prevalence rate as low as a 0,02%-0,1% (i.e. between 2 virus shedders per 10000 and 1 virus shedder per 1000). The wastewater testing method was used to monitor a Danish Community after a COVID-19 outbreak and it was shown that the method can be used as a semi-quantitative method to monitor the development of an outbreak.</jats:p>

Protocol for Safe, Affordable, and Reproducible Isolation and Quantitation of SARS-CoV-2 RNA from Wastewater

“…This result should ameliorate concerns about longer term storage of wastewater samples if they cannot be processed immediately. In our hands, SARS-CoV-2 quantitation was not impaired in the short term by storage at 4 °C either with or without PEG/NaCl, an outcome similarly reflected by other studies [18, 19]. However, storing the pasteurized samples at 4 □ for 72 hours without added PEG and NaCl negatively impacted the recovery of N1 copies by RT-qPCR.…”

Wastewater Analysis of SARS-CoV-2 as a Predictive Metric of Positivity Rate for a Major Metropolis

“…We observed a strong cross-correlation up to fourteen days (Spearman r > 0.8 for smoothed datasets and r > 0.6 for raw datasets), suggesting that detection of SARS-CoV-2 RNA in wastewater could serve as a leading indicator of the positivity rate for community recorded infections. Previous studies have reported the detection of SARS-CoV-2 RNA in influent wastewater prior to known clinical cases in sewersheds ( 6, 7, 20, 21 ). Peccia et al measured SARS-CoV-2 RNA in primary settled solids and found that concentrations of SARS-CoV-2 RNA in the solids were 0-2d ahead of SARS-CoV-2 positive test results (by date of specimen collection) ( 16 ).…”

Sewage analysis as a tool for the COVID-19 pandemic response and management: the urgent need for optimised protocols for SARS-CoV-2 detection and quantification

“…The results showed a prevalence of 0.1 case per 100000 when using N1 and N2 targets [ 38 ], while N3 and E targets provided a positive signal at 3.5 cases per 100000 people [ 21 ] [ preprint] . Another study by Jorgensen et al [99][preprint] has estimated that a positive signal of the viral RNA in wastewater may be produced if there are around 3 cases per 10000 people. However, it is highlighted by the authors that this estimation is based on assumptions that may likely convey lower than normally accepted precision.…”