Rapid detection of SARS-CoV-2 with CRISPR-Cas12a
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 sensitiv...
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| creator | Xiong, Dan Dai, Wenjun Gong, Jiaojiao Li, Guande Liu, Nansong Wu, Wei Pan, Jiaqiang Chen, Chen Jiao, Yingzhen Deng, Huina Ye, Junwei Zhang, Xuanxuan Huang, Huiling Li, Qianyun Xue, Liang Zhang, Xiuming Tang, Guanghui |
| description | 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. |
| doi_str_mv | 10.1371/journal.pbio.3000978 |
| format | Article |
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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.</description><identifier>ISSN: 1545-7885</identifier><identifier>ISSN: 1544-9173</identifier><identifier>EISSN: 1545-7885</identifier><identifier>DOI: 10.1371/journal.pbio.3000978</identifier><identifier>PMID: 33320883</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Base Sequence ; Biology and life sciences ; Control ; Coronaviridae ; Coronaviruses ; COVID-19 Testing - methods ; CRISPR ; CRISPR-Cas Systems - genetics ; Deoxyribonucleic acid ; DNA ; DNA-directed RNA polymerase ; Engineering and Technology ; Epidemics ; Fluorescence ; Genomes ; Humans ; Medicine and health sciences ; Methods and Resources ; Middle East respiratory syndrome ; Molecular diagnostic techniques ; Mutation ; Nucleotides ; Pandemics ; Recombinase ; Reproducibility of Results ; Research and Analysis Methods ; Respiration ; Respiratory diseases ; Room temperature ; SARS-CoV-2 - genetics ; SARS-CoV-2 - isolation & purification ; Sensitivity and Specificity ; Severe acute respiratory syndrome ; Severe acute respiratory syndrome coronavirus 2 ; Substrates ; Transcription ; Viral diseases ; Viruses</subject><ispartof>PLoS biology, 2020-12, Vol.18 (12), p.e3000978-e3000978</ispartof><rights>COPYRIGHT 2020 Public Library of Science</rights><rights>2020 Xiong et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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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.</description><subject>Base Sequence</subject><subject>Biology and life sciences</subject><subject>Control</subject><subject>Coronaviridae</subject><subject>Coronaviruses</subject><subject>COVID-19 Testing - methods</subject><subject>CRISPR</subject><subject>CRISPR-Cas Systems - genetics</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA-directed RNA polymerase</subject><subject>Engineering and Technology</subject><subject>Epidemics</subject><subject>Fluorescence</subject><subject>Genomes</subject><subject>Humans</subject><subject>Medicine and health sciences</subject><subject>Methods and Resources</subject><subject>Middle East respiratory syndrome</subject><subject>Molecular diagnostic 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detection of SARS-CoV-2 with CRISPR-Cas12a</title><author>Xiong, Dan ; Dai, Wenjun ; Gong, Jiaojiao ; Li, Guande ; Liu, Nansong ; Wu, Wei ; Pan, Jiaqiang ; Chen, Chen ; Jiao, Yingzhen ; Deng, Huina ; Ye, Junwei ; Zhang, Xuanxuan ; Huang, Huiling ; Li, Qianyun ; Xue, Liang ; Zhang, Xiuming ; Tang, Guanghui</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c676t-c84cdb6b953e2cef6ae1a1872d5b2dac513fde137c04ccbd17325414b9ba1bf73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Base Sequence</topic><topic>Biology and life sciences</topic><topic>Control</topic><topic>Coronaviridae</topic><topic>Coronaviruses</topic><topic>COVID-19 Testing - methods</topic><topic>CRISPR</topic><topic>CRISPR-Cas Systems - genetics</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA-directed RNA polymerase</topic><topic>Engineering and 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Biol</addtitle><date>2020-12-15</date><risdate>2020</risdate><volume>18</volume><issue>12</issue><spage>e3000978</spage><epage>e3000978</epage><pages>e3000978-e3000978</pages><issn>1545-7885</issn><issn>1544-9173</issn><eissn>1545-7885</eissn><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.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>33320883</pmid><doi>10.1371/journal.pbio.3000978</doi><orcidid>https://orcid.org/0000-0003-4585-9161</orcidid><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central |
| subjects | Base Sequence Biology and life sciences Control Coronaviridae Coronaviruses COVID-19 Testing - methods CRISPR CRISPR-Cas Systems - genetics Deoxyribonucleic acid DNA DNA-directed RNA polymerase Engineering and Technology Epidemics Fluorescence Genomes Humans Medicine and health sciences Methods and Resources Middle East respiratory syndrome Molecular diagnostic techniques Mutation Nucleotides Pandemics Recombinase Reproducibility of Results Research and Analysis Methods Respiration Respiratory diseases Room temperature SARS-CoV-2 - genetics SARS-CoV-2 - isolation & purification Sensitivity and Specificity Severe acute respiratory syndrome Severe acute respiratory syndrome coronavirus 2 Substrates Transcription Viral diseases Viruses |
| title | Rapid detection of SARS-CoV-2 with CRISPR-Cas12a |
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