The COVID-19 pandemic has resulted in an urgent need for a rapid, point of care diagnostic testing that could be rapidly scaled on a worldwide level. We developed and tested a highly sensitive and robust assay based on reverse transcription loop mediated isothermal amplification (RT-LAMP) that uses readily available reagents and a simple heat block using contrived spike-in and actual clinical samples. RT-LAMP testing on RNA-spiked samples showed a limit of detection (LoD) of 2.5 copies/μl of viral transport media. RT-LAMP testing directly on clinical nasopharyngeal swab samples in viral transport media had an 85% positive percentage agreement (PPA) (17/20), and 100% negative percentage agreement (NPV) and delivered results in 30 min. Our optimized RT-LAMP based testing method is a scalable system that is sufficiently sensitive and robust to test for SARS-CoV-2 directly on clinical nasopharyngeal swab samples in viral transport media in 30 min at the point of care without the need for specialized or proprietary equipment or reagents. This cost-effective and efficient one-step testing method can be readily available for COVID-19 testing world-wide, especially in resource poor settings.
【저자키워드】 viral infection, Diagnostic markers, 【초록키워드】 SARS-CoV-2, equipment, COVID-19 pandemic, point of care, diagnostic, Nasopharyngeal swab, clinical samples, RT-LAMP, COVID-19 testing, Viral, Transport media, limit of detection, diagnostic testing, reverse transcription, isothermal amplification, viral transport media, resource, reagents, positive, reagent, nasopharyngeal swab sample, robust, tested, in viral, NPV, scaled, 【제목키워드】 SARS-CoV-2, diagnostic, RNA extraction, diagnostic testing, Direct,