Monitoring COVID-19 In Wastewater With RNA Sequencing

Coronavirus disease 2019, or more commonly known as COVID-19 has brought the world to halt ever since it started spreading. This ongoing pandemic has caused a global emergency and is identified through symptoms like fatigue, body pain, dry cough, fever, and in serious cases, loss of smell and taste. This illness caused by serious acute respiratory syndrome coronavirus 2 (SARS-CoV-2) initiated from China at the end of the year 2019. This was declared as a global pandemic by World Health Organization (WHO) on March 11th 2020 because the disease was spreading like wildfire. 

There have been millions of cases reported from all parts of the world, with almost all the countries experiencing lockdowns along with contact restrictions in order to prevent the disease from spreading. 

In the past few months, wastewater epidemiology (WBE) has come out as a promising method to give early warning of disease outbreaks. It has also offered vital information for the public for patients that are asymptotic. Research and studies on the disease and its symptoms are ongoing and some recent studied have suggested that the detection of SARS-CoV-2 in feces and urine from those who have been tested positive implies that the disease is present in the streams of wastewater treatment plants (WWTPs). 

With the help of environmental surveillance in Wastewater epidemiology (WBE), predicting the whole status of a particular catchment area has become easier than what it would be with clinical surveillance. WBE can be of great use in outbreaks in the overall area, especially by testing the wastewater sample over time. On the other hand, clinical surveillance needs more effort, time, and cost to collect sample and then test it. Moreover, another big advantage of WBE is that it captures people that have asymptomatic and pre-symptomatic infections that are not a part of clinical surveillance. Various researches have shown that wastewater monitoring is able to identify outbreaks of poliovirus and norovirus way before the clinical surverys. (Hovi 2012).

The early studies on the subject reported detection of SARS-CoV-2 RNA in wastewater in several countries like Italy, Australia, and the USA. One of the preliminary studies on the surveillance of COVID-19 in wastewater was done in Australia. In this research, SARS-CoV-2 was detected in two samples in a span of six days from the same WWTP, using both sequencing and qPCR. Researchers in Netherlands conducted tests in sewage of six cities along with the airport for SARS-CoV-2 RNA, focusing on the nucleocapsid (N) and the envelope E gene. Results of these tests proved that the sewage samples were found positive for the N gene. Another research group in Italy worked on 12 influent sewage samples from the WWTPs in Milan and Rome. They concluded that 6 samples came out positive out of 12. 

The main goal of this study was to establish a WBE surveillance system for SARS-CoV-2 in Frankfurt, Germany. This data is to be further used as warning systems in the coming years. WWTP data can easily add valuable information and also assist in decision making on many public and societal restrictions. It can also aid in easing virus concentration. (Agrawal, Orschler and Lackner 2021)

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