Wastewater Monitoring with Digital PCR

Wastewater Monitoring and Surveillance using digital PCR

Digital PCR Enables Breakthroughs in Quantitation and Monitoring

Screening wastewater, or sewage, for the presence of SARS-CoV-2 viral RNA can be an effective monitoring method. Wastewater monitoring enables broad collection of surveillance data – even in areas that have limited access to healthcare or testing facilities. Because natural sewage is highly heterogeneous, a method capable of identifying very rare target RNA from a mixture of non-target nucleic acid molecules is required.

While quantitative PCR (qPCR) is the current standard for COVID-19 clinical testing, the resulting data can be highly variable due to inadequate sample dilution or chemical contamination.1,2 The Absolute Q Digital PCR (dPCR) platform is better suited for detecting SARS-CoV-2 targets from wastewater samples because it overcomes the problem of variability, reduces the impact of many PCR inhibitors, and eliminates the need for standard curves.

Fight Outbreaks With Early Warning

Viral RNA has been detected in environmental samples before symptoms appear.3 The Absolute Q facilitates accurate and sensitive detection of these rare molecules.

In this webinar, our panelists of experts shared their thoughts on how the data collected from SARS-CoV-2 wastewater surveillance efforts are being used and contextualized, as well as the role wastewater-based epidemiology could play in communities after COVID-19 vaccination.

Wastewater Monitoring for SARS-CoV-2 Using the Absolute Q

In this study, we demonstrate SARS-CoV-2 detection and quantification alongside the normalization controls for four wastewater samples collected by the University of Arizona WEST center during their wastewater epidemiology testing efforts.

Tech Note: Detect SARS-CoV-2 alongside human fecal and process controls

This tech note demonstrates the performance of Combinati’s SARS-CoV-2 Wastewater Surveillance 4-plex kit. The assay was designed to detect the N1 and N2 SARS-CoV-2 viral RNA targets alongside the human fecal control, Pepper Mild Mottle virus (PMMoV). In addition to these three targets, the assay also integrates the process control Bovine Coronavirus (BCoV). This inactivated virus, which is not generally present in community sewer systems, is spiked into the initial raw sewage sample before downstream processing. Its similarity to human SARS-CoV-2 allows it to be used as a surrogate to monitor the overall efficiency of sample processing.

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References

  1. Sean C. Taylor et al. Droplet Digital PCR versus qPCR for gene expression analysis with low abundant targets: from variable nonsense to publication quality data. Scientific Reports. 2017 May 25;7(1):2409.
  2. Dong, Lianhua, et al. “Highly Accurate and Sensitive Diagnostic Detection of SARS-CoV-2 by Digital PCR.” MedRxiv, Cold Spring Harbor Laboratory Press, 1 Jan. 2020.
  3. Peccia, Jordan, et al. “SARS-CoV-2 RNA Concentrations in Primary Municipal Sewage Sludge as a Leading Indicator of COVID-19 Outbreak Dynamics.” MedRxiv, Cold Spring Harbor Laboratory Press, 1 Jan. 2020, doi.org/10.1101/2020.05.19.20105999.