tetano
Editor, Senior Moderator
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[SUP] 1 [/SUP], Kun Yin[SUP] 1 [/SUP], Ziyue Li[SUP] 1 [/SUP], Maroun M Sfeir[SUP] 2 [/SUP], Changchun Liu[SUP] 3 [/SUP]
Affiliations
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.
. 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[SUP] 1 [/SUP], Kun Yin[SUP] 1 [/SUP], Ziyue Li[SUP] 1 [/SUP], Maroun M Sfeir[SUP] 2 [/SUP], Changchun Liu[SUP] 3 [/SUP]
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.