A CRISPR-Cas12a-based specific enhancer for more sensitive detection of SARS-CoV-2 infection
Abstract
High Ct-values falling in the grey zone are frequently encountered in SARS-CoV-2 detection by real-time reverse transcription PCR (rRT-PCR) and have brought urgent challenges in diagnosis of samples with low viral load. Based on the single-stranded DNA reporter trans-cleavage activity by Cas12a upon target DNA recognition, we create a <underline>S</underline> pecific <underline>E</underline> nhancer for detection of PCR-amplified <underline>N</underline> ucleic <underline>A</underline> cids (SENA) to confirm SARS-CoV-2 detection through specifically targeting its rRT-PCR amplicons. SENA is highly sensitive, with its limit of detection being at least 2 copies/reaction lower than that of the corresponding rRT-PCR, and highly specific, which identifies both false-negative and false-positive cases in clinic applications. SENA provides effective confirmation for nucleic acid amplification-based molecular diagnosis, and may immediately eliminate the uncertainty problems of rRT-PCR in SARS-CoV-2 clinic detection.
One Sentence Summary
CRISPR-Cas12a-based COVID-19 diagnosis.
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