Multiplexed detection of SARS-CoV-2 and other respiratory infections in high throughput by SARSeq.

Yelagandula R, Bykov A, Vogt A, Heinen R, Özkan E, Strobl MM, Baar JC, Uzunova K, Hajdusits B, Kordic D, Suljic E, Kurtovic-Kozaric A, Izetbegovic S, Schaeffer J, Hufnagl P, Zoufaly A, Seitz T, VCDI , Födinger M, Allerberger F, Stark A, Cochella L, Elling U

Nat Commun 12 (1) 3132 [2021-05-25; online 2021-05-25]

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.

Category: Genomics & transcriptomics

Category: Other

Type: Journal article

PubMed 34035246

DOI 10.1038/s41467-021-22664-5

Crossref 10.1038/s41467-021-22664-5

pii: 10.1038/s41467-021-22664-5
GEO: GSE163688 Sequencing data for all experiments
Primer sequences

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