The recent outbreak of human infections caused by SARS-CoV-2, the third zoonotic coronavirus has raised great public health concern globally. Rapid and accurate diagnosis of this novel pathogen posts great challenges not only clinically but also technologically. Metagenomic next-generation sequencing (mNGS) and reverse-transcription PCR (RT-PCR) have been the most commonly used molecular methodologies. However, each has their own limitations. In this study, we developed an isothermal, CRISPR-based diagnostic for COVID-19 with near single-copy sensitivity. The diagnostic performances of all three technology platforms were also compared. Our study aimed to provide more insights into the molecular detection of SARS-CoV-2, and also to present a novel diagnostic option for this new emerging virus. Author summary The recently discovered betacoronavirus severe acute respiratory syndrome (SARS)-CoV-2 has caused an outbreak of Coronavirus Disease 2019 (COVID-19) that rapidly developed into a global pandemic. The surging demand for rapid screening and identification of COVID-19 posts great diagnostic challenges. A lack of rapid and accurate molecular tools has hampered efficient public health responses to the viral threat. Here, we harnessed the unique collateral activity of programmable CRISPR/Cas13a to develop CRISPR-COVID, a rapid and sensitive diagnostic for SARS-CoV-2 infection, and compared it to sequencing-based metagenomic and RT-PCR-based assays in a clinical cohort. CRISPR-COVID demonstrated a sensitivity level of near single copy and was highly specific without cross reacting to related pathogens. CRISPR-COVID takes only 40 mins and requires no sophiscated thermo-cyclers, providing a valuable alternative to the conventional RT–PCR assay to circumvent the bottlenecks in assay turnaround time, equipment and reagent supplies for COVID-19 testing.
【초록키워드】 COVID-19, coronavirus disease, public health, SARS-CoV-2, Coronavirus disease 2019, coronavirus, equipment, SARS-COV-2 infection, Sequencing, diagnostic, Diagnosis, RT-PCR, virus, Betacoronavirus, global pandemic, CRISPR, sensitivity, Cohort, COVID-19 testing, Viral, PCR, pathogen, outbreak, Next-generation sequencing, Public health response, Rapid, Pathogens, cross, molecular, respiratory, platform, isothermal, acute respiratory syndrome, human infection, human infections, Zoonotic coronavirus, bottlenecks, single copy, RT–PCR, limitations, lack, develop, caused, clinically, raised, unique, demonstrated, circumvent, bottleneck, metagenomic, 【제목키워드】 diagnostic, development,