SARS-CoV-2 RNA was detected using previously described primer and probe sequences targeting the E gene.
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Briefly, Total nucleic acids were extracted from 500 μL VTM on the QIAsymphony SP using the QIAsymphony DSP Virus/Pathogen Midi Kit (both from Qiagen, Germantown, MD) according to the manufacturer’s recommendations, and eluted in 60 μL buffer AVE. The SHC test was performed as described in the EUA documentation. Clinical specimens and reference testingĮighty nasopharyngeal swab samples in viral transport medium (VTM) collected between March 18 and March 28, 2020, from patients under investigation for SARS-CoV-2 submitted to the Stanford Health Care Clinical Virology Laboratory for diagnostic testing, were included in this study. Given the need to rapidly deploy SARS-CoV-2 diagnostic tests in response to the COVID-19 pandemic, these data are important to inform laboratory decision-making and to guide clinical management.ģ.2. To evaluate this isothermal method, as well as two additional rRT-PCR assays, the N1 and N2 components of the US Centers for Disease Control and Prevention (CDC) assay and a commercial multiplex assay targeting the envelope ( E) and spike ( S) genes (Altona Diagnostics), we tested nasopharyngeal samples from individuals under investigation for COVID-19 using the Stanford Health Care (SHC) Clinical Virology Laboratory EUA assay as reference. One such isothermal assay is the Atila iAMP COVID-19 detection kit, which targets the open reading frame 1ab (ORF1ab) region and nucleocapsid ( N) gene in a single channel and utilizes the detection RNase P in a separate channel as the internal control. ,, , ] There are, however, a limited number of isothermal amplification methods that have been developed for the detection of SARS-CoV-2 RNA.
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The current standard of care for diagnosis of active infection is the detection of viral RNA from respiratory specimens by real-time, reverse transcription polymerase chain reaction (rRT-PCR).
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Accurate diagnostics for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are critical to identifying and managing individuals with Coronavirus Disease-2019 (COVID-19).