A multiplexed, next generation sequencing platform for high-throughput detection of SARS-CoV-2
Marie-Ming Aynaud, J. Javier Hernández, Seda Barutcu, Ulrich Braunschweig, Kin Chan, Joel D. Pearson, Daniel Trcka, Suzanna L. Prosser, Jaeyoun Kim, Miriam Barrios‐Rodiles, Mark Jen, Siyuan Song, Jess Shen, Christine Bruce, Bryn Hazlett, Susan M. Poutanen, Liliana Attisano, Rod Bremner, Benjamin J. Blencowe, Tony Mazzulli, Hong Han, Laurence Pelletier, Jeffrey L. Wrana
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.