Detection of SARS-CoV-2 in wastewater in Japan by multiple molecular assays-implication for wastewater-based epidemiology (WBE)

Abstract

Presence of SARS-coronavirus-2 (SARS-CoV-2) in wastewater sample has been documented in several countries. Wastewater-based epidemiology (WBE) is potentially effective for early warning of COVID-19 outbreak. The purpose of this study was to verify the detection limit of WBE for COVID-19. In total, 27 influent wastewater samples were collected from four wastewater treatment plants in Ishikawa and Toyama prefectures in Japan. During the study period, numbers of the confirmed COVID-19 cases in these prefectures increased from almost 0 to around 20 per 100,000 peoples. SARS-CoV-2 RNA in the samples were identified by several PCR-based assays. Among the 27 samples, 7 were positive for SARS-CoV-2 by at least one out of the three quantitative RT-PCR assays. These samples were also positive by RT-nested PCR assays. The detection frequency became higher when the number of total confirmed SARS-CoV-2 cases in 100,000 peoples became above 10 in each prefecture. However, SARS-CoV-2 could also be detected with a low frequency when the number was below 1.0. Considering that the number of the confirmed cases does not necessarily reflect the actual prevalence of the infection at the time point, data on the relationship between the number of infection cases and concentration in wastewater needs to be accumulated further.

A simplified approach to monitoring the COVID-19 epidemiologic situation using waste water analysis and its application in Russia

“…The copyright holder for this this version posted September 23, 2020. ; https://doi.org/10.1101/2020.09.21.20197244 doi: medRxiv preprint between 10 2 -10 4 copies•l -1 , and in feces from -10 10 copies•g -1 (Wu et al, 2020;Wurtzer et al, 2020;Wölfel et al, 2020;La Rosa et al, 2020;Farkas et al, 2020;Haramoto et al, 2020;Bhowmick et al, 2020). Taking into account the reported volume of wastewater produced by citizens (from to l), it can be calculated that the lowest detectable infection rate using WBE methods ranges between 10 -1 and 10 -4 % (Hata et al;Wurtzer et al, 2020;Ahmed et al, 2020).…”

“…There are additional limitations especially for SARS-CoV-2 viral particle determination, since this virus is an RNA-based one, and therefore RT reaction is included in the determination procedure, and the probability of RT efficiency may also differ depending on many factors. Thus, it has been shown that the detection efficiency of SARS-CoV-2 viral particles ranges from 8.5 to 71.6% depending on the concentration method and the volume of the concentrated sample (Hata et al;Medema et al, 2020;Haramoto et al, 2020;. The results obtained may be dependent on molecular process control that can strongly vary in different lab protocols and influence the final results.…”

“…The results obtained may be dependent on molecular process control that can strongly vary in different lab protocols and influence the final results. As such, a control of other coronaviruses as well as mouse hepatitis virus, coliphage MS2 (ATCC 15597-B1), tobacco mosaic virus, Pseudomonas phage Ф 6 and Murine norovirus have been used in different studies (Hata et al;Medema et al, 2020;Haramoto et al, 2020;. The complexity of the wastewater matrix also limits the detection accuracy SARS-CoV-2.…”

Making Waves Perspectives of Modelling and Monitoring of SARS-CoV-2 in Aquatic Environment for COVID-19 Pandemic

“…Hence, identification and quantification of the viral genome in wastewater can be a reliable indicator of disease prevalence among the communities, as revealed for other similar viruses [46][47][48]. Very recently, studies report detections of RNA of SARS-CoV-2 in wastewater [27,46,[49][50][51][52][53], with detected RNA amount being higher than that expected from reported infection cases. Moreover, SARS-CoV-2 was detected in wastewater 7-10 days earlier than clinically reported cases [46,53,54].…”

“…Very recently, studies report detections of RNA of SARS-CoV-2 in wastewater [27,46,[49][50][51][52][53], with detected RNA amount being higher than that expected from reported infection cases. Moreover, SARS-CoV-2 was detected in wastewater 7-10 days earlier than clinically reported cases [46,53,54]. In summary, wastewater analysis is recommended as a non-invasive initial-warning tool that can help in monitoring the trend and status of COVID-19 spread and as a device for tweaking public health response [35,55,56].…”