tetano
Editor, Senior Moderator
ACS Chem Biol
. 2021 Feb 15.
doi: 10.1021/acschembio.0c00840. Online ahead of print.
Development of a Broadly Applicable Cas12a-Linked Beam Unlocking Reaction for Sensitive and Specific Detection of Respiratory Pathogens Including SARS-CoV-2
Yu Wang[SUP] 1 2 [/SUP], Donglan Liu[SUP] 3 [/SUP], Huihuang Lin[SUP] 4 [/SUP], Dingbin Chen[SUP] 3 [/SUP], Jing Sun[SUP] 3 [/SUP], Yusang Xie[SUP] 4 [/SUP], Xinjie Wang[SUP] 1 [/SUP], Peixiang Ma[SUP] 1 [/SUP], Yan Nie[SUP] 1 [/SUP], Hong Mei[SUP] 1 [/SUP], Baihui Zhao[SUP] 5 [/SUP], Xingxu Huang[SUP] 6 [/SUP], Ge Jiang[SUP] 1 [/SUP], Xianxing Jiang[SUP] 2 [/SUP], Jieming Qu[SUP] 4 [/SUP], Jincun Zhao[SUP] 3 [/SUP], Jia Liu[SUP] 1 3 6 [/SUP]
Affiliations
Abstract
The outbreak of novel coronavirus SARS-CoV-2 has caused a worldwide threat to public health. COVID-19 patients with SARS-CoV-2 infection can develop clinical symptoms that are often confused with the infections of other respiratory pathogens. Sensitive and specific detection of SARS-CoV-2 with the ability to discriminate from other viruses is urgently needed for COVID-19 diagnosis. Herein, we streamlined a highly efficient CRISPR-Cas12a-based nucleic acid detection platform, termed Cas12a-linked beam unlocking reaction (CALIBURN). We show that CALIBURN could detect SARS-CoV-2 and other coronaviruses and influenza viruses with little cross-reactivity. Importantly, CALIBURN allowed accurate diagnosis of clinical samples with extremely low viral loads, which is a major obstacle for the clinical applications of existing CRISPR diagnostic platforms. When tested on the specimens from SARS-CoV-2-positive and negative donors, CALIBURN exhibited 73.0% positive and 19.0% presumptive positive rates and 100% specificity. Moreover, unlike existing CRISPR detection methods that were mainly restricted to respiratory specimens, CALIBURN displayed consistent performance across both respiratory and nonrespiratory specimens, suggesting its broad specimen compatibility. Finally, using a mouse model of SARS-CoV-2 infection, we demonstrated that CALIBURN allowed detection of coexisting pathogens without cross-reactivity from a single tissue specimen. Our results suggest that CALIBURN can serve as a versatile platform for the diagnosis of COVID-19 and other respiratory infectious diseases.
. 2021 Feb 15.
doi: 10.1021/acschembio.0c00840. Online ahead of print.
Development of a Broadly Applicable Cas12a-Linked Beam Unlocking Reaction for Sensitive and Specific Detection of Respiratory Pathogens Including SARS-CoV-2
Yu Wang[SUP] 1 2 [/SUP], Donglan Liu[SUP] 3 [/SUP], Huihuang Lin[SUP] 4 [/SUP], Dingbin Chen[SUP] 3 [/SUP], Jing Sun[SUP] 3 [/SUP], Yusang Xie[SUP] 4 [/SUP], Xinjie Wang[SUP] 1 [/SUP], Peixiang Ma[SUP] 1 [/SUP], Yan Nie[SUP] 1 [/SUP], Hong Mei[SUP] 1 [/SUP], Baihui Zhao[SUP] 5 [/SUP], Xingxu Huang[SUP] 6 [/SUP], Ge Jiang[SUP] 1 [/SUP], Xianxing Jiang[SUP] 2 [/SUP], Jieming Qu[SUP] 4 [/SUP], Jincun Zhao[SUP] 3 [/SUP], Jia Liu[SUP] 1 3 6 [/SUP]
Affiliations
- PMID: 33586431
- DOI: 10.1021/acschembio.0c00840
Abstract
The outbreak of novel coronavirus SARS-CoV-2 has caused a worldwide threat to public health. COVID-19 patients with SARS-CoV-2 infection can develop clinical symptoms that are often confused with the infections of other respiratory pathogens. Sensitive and specific detection of SARS-CoV-2 with the ability to discriminate from other viruses is urgently needed for COVID-19 diagnosis. Herein, we streamlined a highly efficient CRISPR-Cas12a-based nucleic acid detection platform, termed Cas12a-linked beam unlocking reaction (CALIBURN). We show that CALIBURN could detect SARS-CoV-2 and other coronaviruses and influenza viruses with little cross-reactivity. Importantly, CALIBURN allowed accurate diagnosis of clinical samples with extremely low viral loads, which is a major obstacle for the clinical applications of existing CRISPR diagnostic platforms. When tested on the specimens from SARS-CoV-2-positive and negative donors, CALIBURN exhibited 73.0% positive and 19.0% presumptive positive rates and 100% specificity. Moreover, unlike existing CRISPR detection methods that were mainly restricted to respiratory specimens, CALIBURN displayed consistent performance across both respiratory and nonrespiratory specimens, suggesting its broad specimen compatibility. Finally, using a mouse model of SARS-CoV-2 infection, we demonstrated that CALIBURN allowed detection of coexisting pathogens without cross-reactivity from a single tissue specimen. Our results suggest that CALIBURN can serve as a versatile platform for the diagnosis of COVID-19 and other respiratory infectious diseases.