Presence of SARS-Coronavirus-2 in sewage

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

Two procedures were used to extract RNA from the concentrated sewage samples. The samples of February 5, 6 and 7 and March 4 and 5 were processed using the RNeasy PowerMicrobiome Kit (Qiagen, Hilden, Germany) according to the manufacturers protocol. For practical reasons it was decided to use the magnetic extraction reagents of the Biomerieux Nuclisens kit (Biomerieux, Amersfoort, the Netherlands) in combination with the semi-automated KingFisher mL (Thermo Scientific, Bleiswijk, The Netherlands) purification system to extract RNA from the Centricon concentrates of the samples of March 15 and 16 as previously described. 15 Elution of RNA was done in 100 µl elution buffer for both RNA extraction methods. The volume of sewage used for RNA extraction is shown in Table 1 for each of the samples.