Multiplexed detection of SARS-CoV-2 and other respiratory infections in high throughput by SARSeq
| Autor: | Marcus Martin Strobl, Franz Allerberger, Ezgi Özkan, Amina Kurtovic-Kozaric, Peter Hufnagl, Alexander Stark, Ramesh Yelagandula, Alexander Vogt, Aleksandr Bykov, Kristina Uzunova, Juliane Christina Baar, Manuela Födinger, Erna Suljic, Bence Hajdusits, Sebija Izetbegovic, Darja Kordic, Luisa Cochella, Robert Heinen, Justine Schaeffer, Alexander Zoufaly, Tamara Seitz, Ulrich Elling |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Computer science Science General Physics and Astronomy Computational biology medicine.disease_cause Polymerase Chain Reaction Sensitivity and Specificity General Biochemistry Genetics and Molecular Biology DNA sequencing Article law.invention Diagnosis Differential 03 medical and health sciences Viral Proteins 0302 clinical medicine law High-Throughput Screening Assays medicine Humans 030212 general & internal medicine Saliva Throughput (business) Respiratory Tract Infections Polymerase chain reaction Multidisciplinary Clinical Laboratory Techniques SARS-CoV-2 RNA COVID-19 High-Throughput Nucleotide Sequencing General Chemistry Gold standard (test) Amplicon 030104 developmental biology Viral infection Viruses RNA Viral Rhinovirus PCR-based techniques |
| Zdroj: | Nature Communications, Vol 12, Iss 1, Pp 1-17 (2021) Nature Communications |
| ISSN: | 2041-1723 |
| Popis: | 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. Massively parallel but cost-effective testing is essential to monitor the spread of pathogenic agents. Here the authors present SARSseq, which uses a dual indexing strategy in a multiplexed RT-PCR reaction to diagnose SARS-CoV-2 at scale. |
| Databáze: | OpenAIRE |
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