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. 2021 May 25;12:3132. doi: 10.1038/s41467-021-22664-5

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — A Six 96-well plates filled with gargle in HBSS and inactivated in QuickExtract from our in-house pipeline were spiked with RNA from various respiratory viruses. For SARS-CoV-2, a positive gargle sample with Ct value of 30 was diluted as indicated. RNA for all other viruses was obtained from HEK cells 48 h after infection with the virus. Dilution indicated by voluminometric ratio. SARSeq was performed with six primer pairs in one reaction, namely N1 and N3 for SARS-CoV-2, Influenza A virus, human rhinovirus (HRV), and the ribosome. Influenza A substrains and HRV substrains were distinguished based on amplicon sequence variants. B Analysis of false-positive and false-negative rate of the experiment in panel A. As expected, 1:100,000 dilutions of the SARS-CoV-2 sample with a Ct of 30 were missed. All other positive samples were detected for all viral spike-ins.