Wastewater-based epidemiology has become an important tool for the surveillance of SARS-CoV-2 outbreaks. However, the detection of viruses in sewage is challenging and to date there is no standard method available which has been validated for the sensitive detection of SARS-CoV-2. In this paper, we describe a simple concentration method based on polyethylene glycol (PEG) precipitation, followed by RNA extraction and a one-step quantitative reverse transcription PCR (qRT-PCR) for viral detection in wastewater. PEG-based concentration of viruses is a simple procedure which is not limited by the availability of expensive equipment and has reduced risk of disruption to consumable supply chains. The concentration and RNA extraction steps enable 900–1500× concentration of wastewater samples and sufficiently eliminates the majority of organic matter, which could inhibit the subsequent qRT-PCR assay. Due to the high variation in the physico-chemical properties of wastewater samples, we recommend the use of process control viruses to determine the efficiency of each step. This procedure enables the concentration and the extraction the DNA/RNA of different viruses and hence can be used for the surveillance of different viral targets for the comprehensive assessment of viral diseases in a community.
【저자키워드】 COVID-19, public health, sewage, environmental samples, wastewater virology, 【초록키워드】 SARS-CoV-2, Epidemiology, equipment, qRT-PCR, Variation, virus, RNA extraction, Outbreaks, Surveillance, Community, target, viral disease, Viral detection, reverse transcription PCR, Quantitative, Concentration, Efficiency, glycol, reduced risk, qRT-PCR assay, subsequent, inhibit, can be used, majority, determine, physico-chemical property, 【제목키워드】 qRT-PCR, wastewater, SARS-CoV-2 RNA, polyethylene,