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

Nat Commun. 2021 May 25;12(1):3132. doi: 10.1038/s41467-021-22664-5.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • COVID-19 / diagnosis
  • Clinical Laboratory Techniques*
  • Diagnosis, Differential
  • High-Throughput Nucleotide Sequencing
  • High-Throughput Screening Assays*
  • Humans
  • Polymerase Chain Reaction
  • RNA, Viral / genetics
  • Respiratory Tract Infections / diagnosis*
  • Respiratory Tract Infections / virology
  • SARS-CoV-2 / genetics
  • SARS-CoV-2 / isolation & purification
  • Saliva / virology
  • Sensitivity and Specificity
  • Viral Proteins / genetics
  • Viruses / classification
  • Viruses / genetics
  • Viruses / isolation & purification*

Substances

  • RNA, Viral
  • Viral Proteins