Abstract
Quantifying severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in clinical samples is crucial for early diagnosis and timely medical treatment of coronavirus disease 2019. Here, we describe a digital warm-start CRISPR (dWS-CRISPR) assay for sensitive quantitative detection of SARS-CoV-2 in clinical samples. The dWS-CRISPR assay is initiated at above 50 °C and overcomes undesired premature target amplification at room temperature, enabling accurate and reliable digital quantification of SARS-CoV-2. By targeting SARS-CoV-2’s nucleoprotein gene, the dWS-CRISPR assay is able to detect down to 5 copies/μl SARS-CoV-2 RNA in the chip. It is clinically validated by quantitatively determining 32 clinical swab samples and three clinical saliva samples. Moreover, it has been demonstrated to directly detect SARS-CoV-2 in heat-treated saliva samples without RNA extraction. Thus, the dWS-CRISPR method, as a sensitive and reliable CRISPR assay, facilitates accurate SARS-CoV-2 detection toward digitized quantification.
Keywords: COVID-19; CRISPR-Cas12a; Digital warm-start CRISPR assay; Nucleic acid quantification; SARS-CoV-2 detection.
【저자키워드】 COVID-19, CRISPR-Cas12a, SARS-CoV-2 detection, Digital warm-start CRISPR assay, Nucleic acid quantification, 【초록키워드】 Treatment, coronavirus disease, SARS-CoV-2, Saliva, Coronavirus disease 2019, coronavirus, severe acute respiratory syndrome Coronavirus, RNA extraction, early diagnosis, CRISPR, amplification, clinical samples, SARS-CoV-2 detection, Digital, Premature, temperature, SARS-CoV-2 RNA, quantification, Quantitative, room temperature, acute respiratory syndrome, acute respiratory syndrome coronavirus, acute respiratory syndrome coronavirus 2, ChIP, swab sample, clinical sample, detect, clinically, facilitate, overcome, demonstrated, initiated, 【제목키워드】 SARS-CoV-2, sensitive, Quantitative, clinical sample,