As to date, more than 49 million confirmed cases of Coronavirus Disease 19 (COVID-19) have been reported worldwide. Current diagnostic protocols use qRT-PCR for viral RNA detection, which is expensive and requires sophisticated equipment, trained personnel and previous RNA extraction. For this reason, we need a faster, direct and more versatile detection method for better epidemiological management of the COVID-19 outbreak. In this work, we propose a direct method without RNA extraction, based on the Loop-mediated isothermal amplification (LAMP) and Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR associated protein (CRISPR-Cas12) technique that allows the fast detection of SARS-CoV-2 from patient samples with high sensitivity and specificity. We obtained a limit of detection of 16 copies/μL with high specificity and at an affordable cost. The diagnostic test readout can be done with a real-time PCR thermocycler or with the naked eye in a blue-light transilluminator. Our method has been evaluated on a small set of clinical samples with promising results.
【저자키워드】 COVID-19, SARS-CoV-2, Diagnosis, CRISPR-Cas12a, LAMP, 【초록키워드】 equipment, qRT-PCR, diagnostic, RNA extraction, Protein, specificity, COVID-19 outbreak, Sensitivity and specificity, Real-time PCR, Loop-mediated isothermal amplification, management, Patient, limit of detection, epidemiological, Viral RNA, confirmed case, diagnostic protocol, current, thermocycler, clinical sample, reported, evaluated, faster, Palindromic, 【제목키워드】 detection, RNA, Direct,