Date: 6/09/2020 01:03:03
From: sarahs mum
ID: 1615209
Subject: how do you find Covid in the sewerage system?
Date: 6/09/2020 02:24:32
From: mollwollfumble
ID: 1615223
Subject: re: how do you find Covid in the sewerage system?
sarahs mum said:
Try this article. From the Netherlands.
https://www.medrxiv.org/content/10.1101/2020.03.29.20045880v2.full.pdf
“To determine if SARS-CoV-2 is present in sewage during the emergence of COVID-19 in the Netherlands, sewage samples of 7 cities and the airport were tested using RT-PCR against three fragments of the nucleocapsid protein gene (N1-3) and one fragment of the envelope protein gene (E). The first case was reported in the Netherlands on February 27. On March 5, the N1 fragment was detected in sewage of five sites.”
“Before the onset of the epidemic in the Netherlands, wastewater treatment plants (WWTP) were selected that served 2 large and 3 medium sized cities and the main airport. The operators of each WWTP collected a sample of 250 mL over 24 h.”
“RNA was isolated on the day of sampling. Bacteria were removed from the samples by pelleting using centrifugation at 4654 g for 30 mins. A volume of 100-200 ml supernatant was filtered through Centricon® Plus-70 centrifugal ultrafilter with a cut-off of 100 kDa by centrifugation at 1500 g for 15 minutes. The resulting concentrate was split in two parts for: 1) quantitative culturing for F-specific RNA phages and 2) for RNA extraction and RT-PCR.”
RNA extracted “using the RNeasy PowerMicrobiome Kit, and the magnetic extraction reagents of the Biomerieux Nuclisens kit in combination with the semi-automated KingFisher mL.”
