Abstract
We demonstrate the application of a rapid pre-concentration protocol and a magnetic bead-based RNA extraction method for the detection of SARS-CoV-2 RNA from raw wastewater without the need for extensive sample treatment.
Highlights
Over four million people as of July 2021.1 SARS-CoV-2 is an enveloped, positively charged, single-stranded ribonucleic acid (RNA) virus (60 to 140 nm) of the beta coronavirus genus.[2,3] COVID-19 is characterized as a respiratory illness, many infected patients present with gastrointestinal distress.[4]
In the first of this series of preliminary experiments to assess the increase in sensitivity of the direct RNA extraction method (ESI†), influent 24-h composite wastewater samples were spiked with HI-SCV-2 to 1 × 103 GU mL−1 and analyzed using the magnetic bead-based RNA extraction protocol with and without the preconcentration step in biological triplicates to determine whether RNA detection rate and recovery were higher using the enhanced extraction protocol
These preliminary results demonstrate that the pre-concentration step in combination with the magnetic beads-based RNA extraction method (3/3 detections) was more effective than the direct extraction method (1/3 detections) in detecting spiked viral HI-SCV-2 in the influent wastewater samples
Summary
Over four million people as of July 2021.1 SARS-CoV-2 is an enveloped, positively charged, single-stranded ribonucleic acid (RNA) virus (60 to 140 nm) of the beta coronavirus genus.[2,3] COVID-19 is characterized as a respiratory illness, many infected patients present with gastrointestinal distress.[4]. Surfactant compounds are commonly used in virology to separate viral aggregates and enhance the release of the viral particles attached to solids by reducing the hydrophobic interactions with the adsorbent.[36,37] Recently, the use of a Tween®20-based elution buffer was reported in the elution of SARS-CoV-2 from passive sampling materials.[38] This pre-extraction approach has been adapted to municipal wastewater samples which allows processing of larger volumes, potentially bridging the gap between pre-concentration sample volumes and traditional RNA extraction sample volumes Such advantages allow for greater diversity in monitoring applications, which is valuable for laboratories with limited resources. This study advances the application of magnetic bead extraction approaches through the development of a new method for a rapid and simple extraction of SARS-CoV-2 RNA from whole wastewater samples
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