Biosens Bioelectron . Sensitive quantitative detection of SARS-CoV-2 in clinical samples using digital warm-start CRISPR assay

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Biosens Bioelectron


. 2021 Apr 17;184:113218.
doi: 10.1016/j.bios.2021.113218. Online ahead of print.
Sensitive quantitative detection of SARS-CoV-2 in clinical samples using digital warm-start CRISPR assay


Xiong Ding ¹ , Kun Yin ¹ , Ziyue Li ¹ , Maroun M Sfeir ² , Changchun Liu ³



Affiliations

• PMID: 33878591
• DOI: 10.1016/j.bios.2021.113218

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/?l 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.