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.
【저자키워드】 viral infection, PCR-based techniques, 【초록키워드】 SARS-CoV-2, Saliva, pandemic, COVID-19 pandemic, Sequencing, Infection, diagnostic, RT-PCR, viral spread, virus, diagnostics, Laboratory, RNA, Spread, sensitivity, RNA sequencing, Pathogens, Quantitative, diagnose, Analysis, Amplicon, gold standard, robustness, virions, MONITOR, Human rhinovirus, performed, detect, can be used, unique, demonstrated, other respiratory virus, pathogenic agents, 【제목키워드】 SARS-CoV-2, other respiratory infection,