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Review
. 2022 Aug 8;19(15):9749.
doi: 10.3390/ijerph19159749.

Wastewater Sequencing-An Innovative Method for Variant Monitoring of SARS-CoV-2 in Populations

Michal Tamáš  1   2 , Alena Potocarova  3 , Barbora Konecna  3 , Ľubos Klucar  4 , Tomas Mackulak  1
Affiliations
Review

Wastewater Sequencing-An Innovative Method for Variant Monitoring of SARS-CoV-2 in Populations

Michal Tamáš et al. Int J Environ Res Public Health. .

Abstract

The SARS-CoV-2 outbreak has already affected more than 555 million people, and 6.3 million people have died. Due to its high infectivity, it is crucial to track SARS-CoV-2 outbreaks early to prevent the spread of infection. Wastewater monitoring appears to be a powerful and effective tool for managing epidemiological situations. Due to emerging mutations of SARS-CoV-2, there is a need to monitor mutations in order to control the pandemic. Since the sequencing of randomly chosen individuals is time-consuming and expensive, sequencing of wastewater plays an important role in revealing the dynamics of infection in a population. The sampling method used is a crucial factor and significantly impacts the results. Wastewater can be collected as a grab sample or as a 24 h composite sample. Another essential factor is the sample volume, as is the method of transport used. This review discusses different pretreatment procedures and RNA extraction, which may be performed using various methods, such as column-based extraction, TRIzol, or magnetic extraction. Each of the methods has its advantages and disadvantages, which are described accordingly. RT-qPCR is a procedure that confirms the presence of SARS-CoV-2 genes before sequencing. This review provides an overview of currently used methods for preparing wastewater samples, from sampling to sequencing.

Keywords: COVID-19; RNA extraction; RT-qPCR; mutations; sampling.

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Conflict of interest statement

All authors claim no conflict of interest.

Figures

Figure 1
Figure 1
Scheme describing step-by-step methodologies for SARS-CoV-2 sequencing from wastewater.
Figure 2
Figure 2
Scheme describing the data analysis and variant identification pipeline, with examples of frequently used tools.
See this image and copyright information in PMC

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