A CRISPR-Cas12a-based specific enhancer for more sensitive detection of SARS-CoV-2 infection.

ABSTRACT:

Background

Real-time reverse transcription-PCR (rRT-PCR) has been the most effective and widely implemented diagnostic technology since the beginning of the COVID-19 pandemic. However, fuzzy rRT-PCR readouts with high Ct values are frequently encountered, resulting in uncertainty in diagnosis.

Methods

A Specific Enhancer for PCR-amplified Nucleic Acid (SENA) was developed based on the Cas12a trans-cleavage activity, which is specifically triggered by the rRT-PCR amplicons of the SARS-CoV-2 Orf1ab (O) and N fragments. SENA was first characterized to determine its sensitivity and specificity, using a systematic titration experiment with pure SARS-CoV-2 RNA standards, and was then verified in several hospitals, employing a couple of commercial rRT-PCR kits and testing various c

SUBMITTER: Huang W 

PROVIDER: S-EPMC7544594 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature