期刊
FRONTIERS IN MEDICINE
卷 8, 期 -, 页码 -出版社
FRONTIERS MEDIA SA
DOI: 10.3389/fmed.2021.627679
关键词
COVID-19; SARS-CoV-2; diagnosis; LAMP; CRISPR-Cas12a
资金
- Consejo Nacional de Ciencia y Tecnologia (CONACyT) [FORDECYT 03/2020]
The study introduced a new method for COVID-19 detection, which does not require RNA extraction, utilizes LAMP and CRISPR-Cas12 technology to rapidly and sensitively detect the virus, and is cost-effective. Initial clinical sample testing results also showed promising prospects.
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/mu L 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.
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