A multiplexed, next generation sequencing platform for high-throughput detection of SARS-CoV-2

Nat Commun. 2021 Mar 3;12(1):1405. doi: 10.1038/s41467-021-21653-y.

Abstract

Population scale sweeps of viral pathogens, such as SARS-CoV-2, require high intensity testing for effective management. Here, we describe "Systematic Parallel Analysis of RNA coupled to Sequencing for Covid-19 screening" (C19-SPAR-Seq), a multiplexed, scalable, readily automated platform for SARS-CoV-2 detection that is capable of analyzing tens of thousands of patient samples in a single run. To address strict requirements for control of assay parameters and output demanded by clinical diagnostics, we employ a control-based Precision-Recall and Receiver Operator Characteristics (coPR) analysis to assign run-specific quality control metrics. C19-SPAR-Seq coupled to coPR on a trial cohort of several hundred patients performs with a specificity of 100% and sensitivity of 91% on samples with low viral loads, and a sensitivity of >95% on high viral loads associated with disease onset and peak transmissibility. This study establishes the feasibility of employing C19-SPAR-Seq for the large-scale monitoring of SARS-CoV-2 and other pathogens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • COVID-19 / genetics
  • COVID-19 / immunology
  • COVID-19 / virology
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Molecular Diagnostic Techniques
  • Nucleic Acid Amplification Techniques
  • RNA, Viral / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • SARS-CoV-2 / growth & development*
  • SARS-CoV-2 / pathogenicity*
  • Viral Load

Substances

  • RNA, Viral