Wastewater Tells The Story About COVID-19 Surveillance

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The Corona virus is the topic that is occupying the society and changing the world. Since the outbreak of the pandemic, SARS-CoV-2 has had a major impact on the lives and health of the world's population. Despite many actions such as operational lockdowns and canceling social interaction, it seems still unpredictable. An important task is to monitor the level of infection and virus transmission to predict possible hotspots and getting into the position to act in advance. The rise of numerous variants of concern underline the importance and the need of efficient surveillance measures.

Since viruses can be excreted through feces, wastewater samples could provide information on the level of infection in certain populations as well as the sequence of circulating variants. Several research groups address this correlation in pilot studies: e.g. Los Angeles County Sanitation District (LACSD), RWTH Aachen, University Hospital Frankfurt, TU Darmstadt and more. The benefit of Wastewater Monitoring is the ability to scale up and test institutions, regions and larger areas at affordable pricing. The detection of both, asymptomatic as well as symptomatic individuals allows to determine if viral concentration is surging or retracting. People and institutions can be alerted in real time of potential high-risk areas before clinical case display the need for action.

Exemplary surveillance in California

To generate this data, appropriate robust and sensitive tools are required. With the development of the Zymo Environ Water RNA Kit (R2042), Zymo Research provides the most effective wastewater workflow:

  • Enhanced viral enrichment; get 8x more viral RNA
  • Pathogen inactivation for safe handling
  • PCR inhibitors removed in one spin
  • ≥ 6 µl elution improves limits of detection

The Zymo Environ Water RNA Kit simplifies water surveillance by facilitating viral/microbial enrichment, inactivating pathogens, and improving limits of detection. The use of Water Concentrating Buffer (WCB) promotes the pelleting of viral capsids and protein-associated RNA, increasing SARS-CoV-2 RNA yields by more than 8-fold. The WCB as well as DNA/RNA Shield™ allow for safe handling as they have been validated for inactivation of pathogens including SARS-CoV-2. Finally, the workflow yields highly concentrated, PCR inhibitor-free RNA ready for downstream applications including RT-qPCR, RT-ddPCR and NGS. (2394)

Detect Down to 8 Viral Copies

Improved Limit of Detection (LOD). SARS-CoV-2 RNA was serially diluted from 5000 to 8 viral copies and spiked into SARS-CoV-2-negative proxy wastewater. RNA was extracted (n=2) using the Zymo Environ Water RNA Kit. RT-qPCR targeting the SARS-CoV-2 N1 gene showed the dilution is linear (R2 = 0.9898) and the workflow was able to achieve an LOD of 8-40 copies

Water Concentrating Buffer Inactivates COVID-19
The Water Concentrating Buffer included in the Zymo Environ Water RNA Kit inactivates SARS-CoV-2 for safe sample processing, while preserving the integrity of viral RNA.

Viral inactivation demonstrated by plaque formation assay. Vero-E6 cells were infected with serial dilutions of SARS-CoV-2, strain Muc-IMB-1. Virus was applied without pre-treatment, or incubated for 48 hours with agarose overlay before fixation with formaldehyde. Plaques were visualized with crystal violet staining.

Affordable, Community-Wide COVID-19 Surveillance


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Letzte Änderungen: 12.05.2021