Author: Fuqing Wu; Amy Xiao; Jianbo Zhang; Xiaoqiong Gu; Wei Lin Lee; Kathryn Kauffman; William Hanage; Mariana Matus; Newsha Ghaeli; Noriko Endo; Claire Duvallet; Katya Moniz; Timothy Erickson; Peter Chai; Janelle Thompson; Eric Alm
Title: SARS-CoV-2 titers in wastewater are higher than expected from clinically confirmed cases Document date: 2020_4_7
ID: atnxkvy6_5
Snippet: We next sought to quantify viral titer in sewage, and to establish our viral enrichment protocol using RT-qPCR. We used the US CDC primer/probe sets targeting the N1, N2, and N3 loci of the SARS-CoV-2 nucleocapsid gene to amplify cDNA reverse transcribed from viral RNA (Table 1) . We tested different steps in our viral enrichment process. First, we examined raw (unfiltered) sewage precipitated with polyethylene glycol 8000 (PEG), which recovers b.....
Document: We next sought to quantify viral titer in sewage, and to establish our viral enrichment protocol using RT-qPCR. We used the US CDC primer/probe sets targeting the N1, N2, and N3 loci of the SARS-CoV-2 nucleocapsid gene to amplify cDNA reverse transcribed from viral RNA (Table 1) . We tested different steps in our viral enrichment process. First, we examined raw (unfiltered) sewage precipitated with polyethylene glycol 8000 (PEG), which recovers both bacterial and viral nucleic acids. Next, we looked at samples taken from 0.2 um filtered sewage: we considered both the material collected on the filter and the filtrate. We found the strongest and most consistent results from the PEG-precipitated viral pellet from the 0.2 um filtrate, which was resuspended in Trizol for RNA extraction ( Table 2) .
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