Title Multiplexed detection of SARS-CoV-2 and other respiratory infections in high throughput by SARSeq.
Author Yelagandula, Ramesh; Bykov, Aleksandr; Vogt, Alexander; Heinen, Robert; Ozkan, Ezgi; Strobl, Marcus Martin; Baar, Juliane Christina; Uzunova, Kristina; Hajdusits, Bence; Kordic, Darja; Suljic, Erna; Kurtovic-Kozaric, Amina; Izetbegovic, Sebija; Schaeffer, Justine; Hufnagl, Peter; Zoufaly, Alexander; Seitz, Tamara; Fodinger, Manuela; Allerberger, Franz; Stark, Alexander; Cochella, Luisa; Elling, Ulrich
Journal Nat Commun Publication Year/Month 2021-May
PMID 34035246 PMCID PMC8149640
Affiliation + expend 1.Institute of Molecular Biotechnology of the Austrian Academy of Science (IMBA), Vienna BioCenter (VBC), Vienna, Austria.

The COVID-19 pandemic has demonstrated the need for massively-parallel, cost-effective tests monitoring viral spread. Here we present SARSeq, saliva analysis by RNA sequencing, a method to detect SARS-CoV-2 and other respiratory viruses on tens of thousands of samples in parallel. SARSeq relies on next generation sequencing of multiple amplicons generated in a multiplexed RT-PCR reaction. Two-dimensional, unique dual indexing, using four indices per sample, enables unambiguous and scalable assignment of reads to individual samples. We calibrate SARSeq on SARS-CoV-2 synthetic RNA, virions, and hundreds of human samples of various types. Robustness and sensitivity were virtually identical to quantitative RT-PCR. Double-blinded benchmarking to gold standard quantitative-RT-PCR performed by human diagnostics laboratories confirms this high sensitivity. SARSeq can be used to detect Influenza A and B viruses and human rhinovirus in parallel, and can be expanded for detection of other pathogens. Thus, SARSeq is ideally suited for differential diagnostic of infections during a pandemic.

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