Presence of SARS-Coronavirus-2 in sewage

Gertjan Medema; Leo Heijnen; Goffe Elsinga; Ronald Italiaander; Anke Brouwer

The concentration of F-specific RNA phages was measured according to ISO 10705 in wastewater before and after the purification and concentration steps, to determine the virus recovery. A previously described non-target RNA fragment 18 was added to the lysed sewage concentrates as an internal control (IC). This IC-RNA consists of an in vitro transcribed RNA fragment with a length of 412 bases derived from dengue virus type 2 and was synthesized as previously described. 18 RT-PCR analyses using primers IC-F (5´-ATGACAGCCACTCCTCCG-3´), IC-R (5´-GGAACGAACCAAACAGTCTTC-3´) and probe IC-P (5´-TexasRed-AGCAGAGACCCATTCCCTCAGAGC-BHQ-3´) were used to detect a dengue virus fragment (length: 149 bp) in order to control the performance of RNA-extraction, RT-PCR and to detect the presence of inhibitors. High-resolution automatic electrophoresis was performed on an Agilent 2100 Bioanalyzer using Agilent 1000 DNA Kit (Agilent, Santa Clara, USA) to analyse the length of the PCR-products. All rights reserved. No reuse allowed without permission.