Aptamer-based Cas14a1 biosensor for amplification-free live pathogenic detection
CRISPR-Cas systems have been employed to detect a large variety of pathogenic microorganisms by simply changing the guide RNA sequence. However, these platforms usually rely on nucleic acid extraction and amplification to achieve good sensitivity. Herein, we developed a new platform for the highly s...
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| Veröffentlicht in: | Biosensors & bioelectronics 2022-09, Vol.211, p.114282-114282, Article 114282 |
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| creator | Wei, Yangdao Tao, Zhenzhen Wan, Lu Zong, Chengli Wu, Jiajia Tan, Xiao Wang, Buhua Guo, Zixuan Zhang, Ling Yuan, Haoyu Wang, Peng Yang, Zhiqing Wan, Yi |
| description | CRISPR-Cas systems have been employed to detect a large variety of pathogenic microorganisms by simply changing the guide RNA sequence. However, these platforms usually rely on nucleic acid extraction and amplification to achieve good sensitivity. Herein, we developed a new platform for the highly specific and sensitive detection of live staphylococcus aureus (S. aureus) based on an Aptamer-based Cas14a1 Biosensor (ACasB), without the need for nucleic acid extraction or amplification. First, the S. aureus specific aptamer was hybrid with a blocker DNA. After the live S. aureus was added, the blocker can be released upon bacteria-aptamer binding. Finally, the released blocker can activate Cas14a1 protein by binding with the sgRNA to generate a change of fluorescent intensity. The ACasB indicates high specificity and sensitivity: it can directly distinguish 400 CFU/ml live S. aureus cells. Comparable to qPCR, the Cas14a1-aptamer biosensor can detect S. aureus with 100% accuracy in complex samples. Therefore, this ACasB for the on-site detection of live S. aureus can broaden its applications in food safety and environmental monitoring.
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| doi_str_mv | 10.1016/j.bios.2022.114282 |
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[Display omitted]</description><identifier>ISSN: 0956-5663</identifier><identifier>EISSN: 1873-4235</identifier><identifier>DOI: 10.1016/j.bios.2022.114282</identifier><identifier>PMID: 35597144</identifier><language>eng</language><publisher>England: Elsevier B.V</publisher><subject>Bacteria diagnostic ; CRISPR-Cas ; Staphylococcus aureus</subject><ispartof>Biosensors & bioelectronics, 2022-09, Vol.211, p.114282-114282, Article 114282</ispartof><rights>2022</rights><rights>Copyright © 2022. Published by Elsevier B.V.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c356t-dada201bfb376b40b7b7bcc8f96190bb7b7a836036af2c08359251945c2aac4c3</citedby><cites>FETCH-LOGICAL-c356t-dada201bfb376b40b7b7bcc8f96190bb7b7a836036af2c08359251945c2aac4c3</cites><orcidid>0000-0002-0667-3279 ; 0000-0002-3398-7396</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0956566322003220$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35597144$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wei, Yangdao</creatorcontrib><creatorcontrib>Tao, Zhenzhen</creatorcontrib><creatorcontrib>Wan, Lu</creatorcontrib><creatorcontrib>Zong, Chengli</creatorcontrib><creatorcontrib>Wu, Jiajia</creatorcontrib><creatorcontrib>Tan, Xiao</creatorcontrib><creatorcontrib>Wang, Buhua</creatorcontrib><creatorcontrib>Guo, Zixuan</creatorcontrib><creatorcontrib>Zhang, Ling</creatorcontrib><creatorcontrib>Yuan, Haoyu</creatorcontrib><creatorcontrib>Wang, Peng</creatorcontrib><creatorcontrib>Yang, Zhiqing</creatorcontrib><creatorcontrib>Wan, Yi</creatorcontrib><title>Aptamer-based Cas14a1 biosensor for amplification-free live pathogenic detection</title><title>Biosensors & bioelectronics</title><addtitle>Biosens Bioelectron</addtitle><description>CRISPR-Cas systems have been employed to detect a large variety of pathogenic microorganisms by simply changing the guide RNA sequence. However, these platforms usually rely on nucleic acid extraction and amplification to achieve good sensitivity. Herein, we developed a new platform for the highly specific and sensitive detection of live staphylococcus aureus (S. aureus) based on an Aptamer-based Cas14a1 Biosensor (ACasB), without the need for nucleic acid extraction or amplification. First, the S. aureus specific aptamer was hybrid with a blocker DNA. After the live S. aureus was added, the blocker can be released upon bacteria-aptamer binding. Finally, the released blocker can activate Cas14a1 protein by binding with the sgRNA to generate a change of fluorescent intensity. The ACasB indicates high specificity and sensitivity: it can directly distinguish 400 CFU/ml live S. aureus cells. Comparable to qPCR, the Cas14a1-aptamer biosensor can detect S. aureus with 100% accuracy in complex samples. Therefore, this ACasB for the on-site detection of live S. aureus can broaden its applications in food safety and environmental monitoring.
[Display omitted]</description><subject>Bacteria diagnostic</subject><subject>CRISPR-Cas</subject><subject>Staphylococcus aureus</subject><issn>0956-5663</issn><issn>1873-4235</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><recordid>eNp9kE1LxDAQhoMo7rr6BzxIj15a890WvCyLX7CgBz2HJJ1qln6ZdAX_vSldPcowDMO88zLzIHRJcEYwkTe7zLg-ZBRTmhHCaUGP0JIUOUs5ZeIYLXEpZCqkZAt0FsIOY5yTEp-iBROizAnnS_SyHkbdgk-NDlAlGx0I1ySZjKELvU_qmLodGlc7q0fXd2ntAZLGfUEy6PGjf4fO2aSCEew0PkcntW4CXBzqCr3d371uHtPt88PTZr1NLRNyTCtdaYqJqQ3LpeHY5DGsLepSxhPN1OqCScykrqnFBRMlFaTkwlKtLbdsha5n38H3n3sIo2pdsNA0uoN-HxSVMs8LzqSIUjpLre9D8FCrwbtW-29FsJpIqp2aHlYTSTWTjEtXB_-9aaH6W_lFFwW3swDil18OvArWQWehcj6iUFXv_vP_AYvQhJM</recordid><startdate>20220901</startdate><enddate>20220901</enddate><creator>Wei, Yangdao</creator><creator>Tao, Zhenzhen</creator><creator>Wan, Lu</creator><creator>Zong, Chengli</creator><creator>Wu, Jiajia</creator><creator>Tan, Xiao</creator><creator>Wang, Buhua</creator><creator>Guo, Zixuan</creator><creator>Zhang, Ling</creator><creator>Yuan, Haoyu</creator><creator>Wang, Peng</creator><creator>Yang, Zhiqing</creator><creator>Wan, Yi</creator><general>Elsevier B.V</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-0667-3279</orcidid><orcidid>https://orcid.org/0000-0002-3398-7396</orcidid></search><sort><creationdate>20220901</creationdate><title>Aptamer-based Cas14a1 biosensor for amplification-free live pathogenic detection</title><author>Wei, Yangdao ; Tao, Zhenzhen ; Wan, Lu ; Zong, Chengli ; Wu, Jiajia ; Tan, Xiao ; Wang, Buhua ; Guo, Zixuan ; Zhang, Ling ; Yuan, Haoyu ; Wang, Peng ; Yang, Zhiqing ; Wan, Yi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c356t-dada201bfb376b40b7b7bcc8f96190bb7b7a836036af2c08359251945c2aac4c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Bacteria diagnostic</topic><topic>CRISPR-Cas</topic><topic>Staphylococcus aureus</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wei, Yangdao</creatorcontrib><creatorcontrib>Tao, Zhenzhen</creatorcontrib><creatorcontrib>Wan, Lu</creatorcontrib><creatorcontrib>Zong, Chengli</creatorcontrib><creatorcontrib>Wu, Jiajia</creatorcontrib><creatorcontrib>Tan, Xiao</creatorcontrib><creatorcontrib>Wang, Buhua</creatorcontrib><creatorcontrib>Guo, Zixuan</creatorcontrib><creatorcontrib>Zhang, Ling</creatorcontrib><creatorcontrib>Yuan, Haoyu</creatorcontrib><creatorcontrib>Wang, Peng</creatorcontrib><creatorcontrib>Yang, Zhiqing</creatorcontrib><creatorcontrib>Wan, Yi</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Biosensors & bioelectronics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wei, Yangdao</au><au>Tao, Zhenzhen</au><au>Wan, Lu</au><au>Zong, Chengli</au><au>Wu, Jiajia</au><au>Tan, Xiao</au><au>Wang, Buhua</au><au>Guo, Zixuan</au><au>Zhang, Ling</au><au>Yuan, Haoyu</au><au>Wang, Peng</au><au>Yang, Zhiqing</au><au>Wan, Yi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Aptamer-based Cas14a1 biosensor for amplification-free live pathogenic detection</atitle><jtitle>Biosensors & bioelectronics</jtitle><addtitle>Biosens Bioelectron</addtitle><date>2022-09-01</date><risdate>2022</risdate><volume>211</volume><spage>114282</spage><epage>114282</epage><pages>114282-114282</pages><artnum>114282</artnum><issn>0956-5663</issn><eissn>1873-4235</eissn><abstract>CRISPR-Cas systems have been employed to detect a large variety of pathogenic microorganisms by simply changing the guide RNA sequence. However, these platforms usually rely on nucleic acid extraction and amplification to achieve good sensitivity. Herein, we developed a new platform for the highly specific and sensitive detection of live staphylococcus aureus (S. aureus) based on an Aptamer-based Cas14a1 Biosensor (ACasB), without the need for nucleic acid extraction or amplification. First, the S. aureus specific aptamer was hybrid with a blocker DNA. After the live S. aureus was added, the blocker can be released upon bacteria-aptamer binding. Finally, the released blocker can activate Cas14a1 protein by binding with the sgRNA to generate a change of fluorescent intensity. The ACasB indicates high specificity and sensitivity: it can directly distinguish 400 CFU/ml live S. aureus cells. Comparable to qPCR, the Cas14a1-aptamer biosensor can detect S. aureus with 100% accuracy in complex samples. Therefore, this ACasB for the on-site detection of live S. aureus can broaden its applications in food safety and environmental monitoring.
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| subjects | Bacteria diagnostic CRISPR-Cas Staphylococcus aureus |
| title | Aptamer-based Cas14a1 biosensor for amplification-free live pathogenic detection |
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