tetano
Editor, Senior Moderator
PLoS Biol
. 2020 Dec 15;18(12):e3000978.
doi: 10.1371/journal.pbio.3000978. eCollection 2020 Dec.
Rapid detection of SARS-CoV-2 with CRISPR-Cas12a
Dan Xiong[SUP] 1 [/SUP], Wenjun Dai[SUP] 2 [/SUP], Jiaojiao Gong[SUP] 2 [/SUP], Guande Li[SUP] 2 [/SUP], Nansong Liu[SUP] 2 [/SUP], Wei Wu[SUP] 1 [/SUP], Jiaqiang Pan[SUP] 2 [/SUP], Chen Chen[SUP] 2 [/SUP], Yingzhen Jiao[SUP] 3 [/SUP], Huina Deng[SUP] 2 [/SUP], Junwei Ye[SUP] 2 [/SUP], Xuanxuan Zhang[SUP] 2 [/SUP], Huiling Huang[SUP] 2 [/SUP], Qianyun Li[SUP] 4 [/SUP], Liang Xue[SUP] 2 [/SUP], Xiuming Zhang[SUP] 1 [/SUP], Guanghui Tang[SUP] 2 [/SUP]
Affiliations
Abstract
The recent outbreak of betacoronavirus Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), which is responsible for the Coronavirus Disease 2019 (COVID-19) global pandemic, has created great challenges in viral diagnosis. The existing methods for nucleic acid detection are of high sensitivity and specificity, but the need for complex sample manipulation and expensive machinery slow down the disease detection. Thus, there is an urgent demand to develop a rapid, inexpensive, and sensitive diagnostic test to aid point-of-care viral detection for disease monitoring. In this study, we developed a clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated proteins (Cas) 12a-based diagnostic method that allows the results to be visualized by the naked eye. We also introduced a rapid sample processing method, and when combined with recombinase polymerase amplification (RPA), the sample to result can be achieved in 50 minutes with high sensitivity (1-10 copies per reaction). This accurate and portable detection method holds a great potential for COVID-19 control, especially in areas where specialized equipment is not available.
. 2020 Dec 15;18(12):e3000978.
doi: 10.1371/journal.pbio.3000978. eCollection 2020 Dec.
Rapid detection of SARS-CoV-2 with CRISPR-Cas12a
Dan Xiong[SUP] 1 [/SUP], Wenjun Dai[SUP] 2 [/SUP], Jiaojiao Gong[SUP] 2 [/SUP], Guande Li[SUP] 2 [/SUP], Nansong Liu[SUP] 2 [/SUP], Wei Wu[SUP] 1 [/SUP], Jiaqiang Pan[SUP] 2 [/SUP], Chen Chen[SUP] 2 [/SUP], Yingzhen Jiao[SUP] 3 [/SUP], Huina Deng[SUP] 2 [/SUP], Junwei Ye[SUP] 2 [/SUP], Xuanxuan Zhang[SUP] 2 [/SUP], Huiling Huang[SUP] 2 [/SUP], Qianyun Li[SUP] 4 [/SUP], Liang Xue[SUP] 2 [/SUP], Xiuming Zhang[SUP] 1 [/SUP], Guanghui Tang[SUP] 2 [/SUP]
Affiliations
- PMID: 33320883
- DOI: 10.1371/journal.pbio.3000978
Abstract
The recent outbreak of betacoronavirus Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), which is responsible for the Coronavirus Disease 2019 (COVID-19) global pandemic, has created great challenges in viral diagnosis. The existing methods for nucleic acid detection are of high sensitivity and specificity, but the need for complex sample manipulation and expensive machinery slow down the disease detection. Thus, there is an urgent demand to develop a rapid, inexpensive, and sensitive diagnostic test to aid point-of-care viral detection for disease monitoring. In this study, we developed a clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated proteins (Cas) 12a-based diagnostic method that allows the results to be visualized by the naked eye. We also introduced a rapid sample processing method, and when combined with recombinase polymerase amplification (RPA), the sample to result can be achieved in 50 minutes with high sensitivity (1-10 copies per reaction). This accurate and portable detection method holds a great potential for COVID-19 control, especially in areas where specialized equipment is not available.