Abstract
The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems have recently received notable attention for their applications in nucleic acid detection. Despite many attempts, the majority of current CRISPR-based biosensors in infectious respiratory disease diagnostic applications still require target preamplifications. This study reports a new biosensor for amplification-free nucleic acid detection via harnessing the trans-cleavage mechanism of Cas13a and ultrasensitive graphene field-effect transistors (gFETs). CRISPR Cas13a-gFET achieves the detection of SARS-CoV-2 and respiratory syncytial virus (RSV) genome down to 1 attomolar without target preamplifications. Additionally, we validate the detection performance using clinical SARS-CoV-2 samples, including those with low viral loads (Ct value >30). Overall, these findings establish our CRISPR Cas13a-gFET among the most sensitive amplification-free nucleic acid diagnostic platforms to date.
Keywords: Amplification-Free Detection; Biosensors; CRISPR Cas13a; Graphene Field-Effect Transistors; SARS-CoV-2.
【저자키워드】 SARS-CoV-2., biosensors, Amplification-Free Detection, CRISPR Cas13a, Graphene Field-Effect Transistors, 【초록키워드】 SARS-CoV-2, Genome, diagnostic, CRISPR, amplification, nucleic acid, Nucleic acid detection, Viral load, respiratory syncytial virus, RSV, Respiratory disease, Ct value, platform, mechanism, Graphene, clustered regularly interspaced short palindromic repeats, majority, notable, palindromic repeat, 【제목키워드】 SARS-CoV-2, detection, respiratory, transistor,