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Updated: Nov 25, 2025

Quantification and Whole Genome Characterization of SARS-CoV-2 RNA in Wastewater and Air Samples
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Comparing analytical methods to detect SARS-CoV-2 in wastewater.

Alba Pérez-Cataluña1, Enric Cuevas-Ferrando1, Walter Randazzo2

  • 1Department of Preservation and Food Safety Technologies, Institute of Agrochemistry and Food Technology, IATA-CSIC, Av. Agustín Escardino 7, Paterna, 46980, Valencia, Spain.

The Science of the Total Environment
|December 18, 2020
PubMed
Summary

Wastewater epidemiology (WBE) for COVID-19 surveillance requires validated methods. This study found that aluminum precipitation combined with automated RNA extraction offers acceptable sensitivity for SARS-CoV-2 detection in wastewater.

Keywords:
Aluminum-based adsorption-precipitationPolyethylene glycol precipitationPorcine epidemic diarrhea virusRT-qPCRSARS-CoV-2Wastewater based epidemiology

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Area of Science:

  • Environmental science
  • Virology
  • Public health

Background:

  • Wastewater-based epidemiology (WBE) is crucial for monitoring infectious diseases like COVID-19.
  • Standardized and validated protocols are needed for reliable SARS-CoV-2 detection in wastewater.

Purpose of the Study:

  • To evaluate and compare different concentration and RNA extraction methods for SARS-CoV-2 detection in wastewater.
  • To benchmark the performance of various protocols for WBE surveillance.

Main Methods:

  • Compared aluminum precipitation and PEG precipitation for concentrating inactivated SARS-CoV-2 and surrogates (PEDV, MgV).
  • Assessed manual spin column and automated magnetic bead-based RNA extraction methods.
  • Determined recovery rates and limits of detection (LoD) for SARS-CoV-2 genomic targets.

Main Results:

  • Aluminum precipitation and automated extraction showed good recovery for SARS-CoV-2 (42.9%) and acceptable LoD values (1.56 log ge/mL for N1).
  • Extraction methods significantly impacted PEDV recovery; concentration methods did not affect SARS-CoV-2 or MgV recovery.
  • High positivity rates for SARS-CoV-2 targets (N1, N2, E, IP2, IP4) were observed in naturally contaminated wastewater samples.

Conclusions:

  • The combination of aluminum precipitation and automated nucleic acid extraction is a sensitive and reliable method for SARS-CoV-2 WBE surveillance.
  • Standardized and validated methods are essential for accurate and consistent WBE data.
  • This approach can provide timely results for public health decision-making.