Simpler and faster Covid-19 testing: Strategies to streamline SARS-CoV-2 molecular assays

• DOI: 10.1016/j.ebiom.2021.103236
• Publication Date: 2021-02-12T04:01:02Z
• Authors: Nuttada Panpradist; Qin Wang; Parker S. Ruth; Jack H. Kotnik; Amy K. Oreskovic; Abraham Miller; Samuel W.A. Stewart; Justin Vrana; Peter D. Han; Ingrid A. Beck; Lea M. Starita; Lisa M. Frenkel; Barry R. Lutz
• Keywords: Gold standard (test); Point-of-Care Testing

BackgroundDetection of SARS-CoV-2 infections is important for treatment, isolation infected and exposed individuals, contact tracing. RT-qPCR the "gold-standard" method to sensitively detect RNA, but most laboratory-developed assays involve complex steps. Here, we aimed simplify by streamlining reaction setup, eliminating RNA extraction, proposing reduced-cost detection workflows that avoid need expensive qPCR instruments.MethodA low-cost RT-PCR based "kit" was developed faster turnaround than CDC protocol. We demonstrated three workflows: two can be deployed in laboratories conducting variable complexity, one could simple enough point-of-care. Analytical sensitivity assessed using spiked simulated nasal matrix. Clinical performance evaluated contrived human matrix (n = 41) clinical specimens collected from individuals with respiratory symptoms 110).FindingThe analytical lyophilised 10 copies/reaction purified 20 when direct lysate Evaluation assay on showed 96.7–100% specificity 100% at ≥20 copies. A head-to-head comparison standard protocol 83.8–94.6% 96.8–100% specificity. found 3.6% indeterminate samples (undetected control), lower 8.1% protocol.InterpretationThis preliminary work should support or commercial entities develop expand access Covid-19 testing. Software guidance development this ongoing enable implementation other settings.FundUSA NIH R01AI140845 Seattle Children's Research Institute