Cascaded signal amplification strategy for ultra-specific, ultra-sensitive, and visual detection of Shigella flexneri
Pathogen infections including Shigella flexneri have posed a significant threat to human health for numerous years. Although culturing and qPCR were the gold standards for pathogen detection, time-consuming and instrument-dependent restrict their application in rapid diagnosis and economically less-...
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| Veröffentlicht in: | Mikrochimica acta (1966) 2024-05, Vol.191 (5), p.271-271, Article 271 |
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| container_title | Mikrochimica acta (1966) |
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| creator | Shi, Yaoqiang Tan, Qi Gong, Tao Li, Qing-yuan Zhu, Ya Duan, Xiaoqiong Yang, Chunhui Ding, Jia-wei Li, Shilin Xie, He Li, Yujia Chen, Limin |
| description | Pathogen infections including
Shigella flexneri
have posed a significant threat to human health for numerous years. Although culturing and qPCR were the gold standards for pathogen detection, time-consuming and instrument-dependent restrict their application in rapid diagnosis and economically less-developed regions. Thus, it is urgently needed to develop rapid, simple, sensitive, accurate, and low-cost detection methods for pathogen detection. In this study, an immunomagnetic beads-recombinase polymerase amplification-CRISPR/Cas12a (IMB-RPA-CRISPR/Cas12a) method was built based on a cascaded signal amplification strategy for ultra-specific, ultra-sensitive, and visual detection of
S. flexneri
in the laboratory. Firstly,
S. flexneri
was specifically captured and enriched by IMB (
Shigella
antibody-coated magnetic beads), and the genomic DNA was released and used as the template in the RPA reaction. Then, the RPA products were mixed with the pre-loaded CRISPR/Cas12a for fluorescence visualization. The results were observed by naked eyes under LED blue light, with a sensitivity of 5 CFU/mL in a time of 70 min. With no specialized equipment or complicated technical requirements, the IMB-RPA-CRISPR/Cas12a diagnostic method can be used for visual, rapid, and simple detection of
S. flexneri
and can be easily adapted to monitoring other pathogens.
Graphical abstract |
| doi_str_mv | 10.1007/s00604-024-06309-0 |
| format | Article |
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Shigella flexneri
have posed a significant threat to human health for numerous years. Although culturing and qPCR were the gold standards for pathogen detection, time-consuming and instrument-dependent restrict their application in rapid diagnosis and economically less-developed regions. Thus, it is urgently needed to develop rapid, simple, sensitive, accurate, and low-cost detection methods for pathogen detection. In this study, an immunomagnetic beads-recombinase polymerase amplification-CRISPR/Cas12a (IMB-RPA-CRISPR/Cas12a) method was built based on a cascaded signal amplification strategy for ultra-specific, ultra-sensitive, and visual detection of
S. flexneri
in the laboratory. Firstly,
S. flexneri
was specifically captured and enriched by IMB (
Shigella
antibody-coated magnetic beads), and the genomic DNA was released and used as the template in the RPA reaction. Then, the RPA products were mixed with the pre-loaded CRISPR/Cas12a for fluorescence visualization. The results were observed by naked eyes under LED blue light, with a sensitivity of 5 CFU/mL in a time of 70 min. With no specialized equipment or complicated technical requirements, the IMB-RPA-CRISPR/Cas12a diagnostic method can be used for visual, rapid, and simple detection of
S. flexneri
and can be easily adapted to monitoring other pathogens.
Graphical abstract</description><identifier>ISSN: 0026-3672</identifier><identifier>EISSN: 1436-5073</identifier><identifier>DOI: 10.1007/s00604-024-06309-0</identifier><identifier>PMID: 38632191</identifier><language>eng</language><publisher>Vienna: Springer Vienna</publisher><subject>Amplification ; Analytical Chemistry ; Antibodies ; Characterization and Evaluation of Materials ; Chemistry ; Chemistry and Materials Science ; CRISPR ; Microengineering ; Nanochemistry ; Nanotechnology ; Original Paper ; Pathogens</subject><ispartof>Mikrochimica acta (1966), 2024-05, Vol.191 (5), p.271-271, Article 271</ispartof><rights>The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.</rights><rights>2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c326t-74163ffb835009d85af92115910def172d31777248720a587ff612d4224d670c3</cites><orcidid>0000-0001-6076-7711</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00604-024-06309-0$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00604-024-06309-0$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27903,27904,41467,42536,51297</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/38632191$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Shi, Yaoqiang</creatorcontrib><creatorcontrib>Tan, Qi</creatorcontrib><creatorcontrib>Gong, Tao</creatorcontrib><creatorcontrib>Li, Qing-yuan</creatorcontrib><creatorcontrib>Zhu, Ya</creatorcontrib><creatorcontrib>Duan, Xiaoqiong</creatorcontrib><creatorcontrib>Yang, Chunhui</creatorcontrib><creatorcontrib>Ding, Jia-wei</creatorcontrib><creatorcontrib>Li, Shilin</creatorcontrib><creatorcontrib>Xie, He</creatorcontrib><creatorcontrib>Li, Yujia</creatorcontrib><creatorcontrib>Chen, Limin</creatorcontrib><title>Cascaded signal amplification strategy for ultra-specific, ultra-sensitive, and visual detection of Shigella flexneri</title><title>Mikrochimica acta (1966)</title><addtitle>Microchim Acta</addtitle><addtitle>Mikrochim Acta</addtitle><description>Pathogen infections including
Shigella flexneri
have posed a significant threat to human health for numerous years. Although culturing and qPCR were the gold standards for pathogen detection, time-consuming and instrument-dependent restrict their application in rapid diagnosis and economically less-developed regions. Thus, it is urgently needed to develop rapid, simple, sensitive, accurate, and low-cost detection methods for pathogen detection. In this study, an immunomagnetic beads-recombinase polymerase amplification-CRISPR/Cas12a (IMB-RPA-CRISPR/Cas12a) method was built based on a cascaded signal amplification strategy for ultra-specific, ultra-sensitive, and visual detection of
S. flexneri
in the laboratory. Firstly,
S. flexneri
was specifically captured and enriched by IMB (
Shigella
antibody-coated magnetic beads), and the genomic DNA was released and used as the template in the RPA reaction. Then, the RPA products were mixed with the pre-loaded CRISPR/Cas12a for fluorescence visualization. The results were observed by naked eyes under LED blue light, with a sensitivity of 5 CFU/mL in a time of 70 min. With no specialized equipment or complicated technical requirements, the IMB-RPA-CRISPR/Cas12a diagnostic method can be used for visual, rapid, and simple detection of
S. flexneri
and can be easily adapted to monitoring other pathogens.
Graphical abstract</description><subject>Amplification</subject><subject>Analytical Chemistry</subject><subject>Antibodies</subject><subject>Characterization and Evaluation of Materials</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>CRISPR</subject><subject>Microengineering</subject><subject>Nanochemistry</subject><subject>Nanotechnology</subject><subject>Original Paper</subject><subject>Pathogens</subject><issn>0026-3672</issn><issn>1436-5073</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><recordid>eNp9kU1P3DAQhq0K1N1u-wd6QJa4cCAw_oidHNGKtkhIHKBny8TjrVfZZLETBP--3s1SJA49WNZonnlszUvIdwYXDEBfJgAFsgCejxJQF_CJzJkUqihBiyMyB-CqEErzGfmS0hqAacXlZzITlRKc1WxOxqVNjXXoaAqrzrbUbrZt8KGxQ-g7moZoB1y9Ut9HOra5KtIWmx1w_lZjl8IQnvGc2s7R55DGrHE4YLNX9J7e_wkrbFtLfYsvHcbwlRx72yb8drgX5PeP64flr-L27ufN8uq2aARXQ6ElU8L7x0qUALWrSutrzlhZM3DomeZOMK01l5XmYMtKe68Yd5Jz6ZSGRizI2eTdxv5pxDSYTUjN7isd9mMyAiTjQmqlMnr6AV33Y8wb2VNQSlZXMlN8oprYpxTRm20MGxtfDQOzC8VMoZgcitmHYiAPnRzU4-MG3b-RtxQyICYg5Va3wvj-9n-0fwEHhZbI</recordid><startdate>20240501</startdate><enddate>20240501</enddate><creator>Shi, Yaoqiang</creator><creator>Tan, Qi</creator><creator>Gong, Tao</creator><creator>Li, Qing-yuan</creator><creator>Zhu, Ya</creator><creator>Duan, Xiaoqiong</creator><creator>Yang, Chunhui</creator><creator>Ding, Jia-wei</creator><creator>Li, Shilin</creator><creator>Xie, He</creator><creator>Li, Yujia</creator><creator>Chen, Limin</creator><general>Springer Vienna</general><general>Springer Nature B.V</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>K9.</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0001-6076-7711</orcidid></search><sort><creationdate>20240501</creationdate><title>Cascaded signal amplification strategy for ultra-specific, ultra-sensitive, and visual detection of Shigella flexneri</title><author>Shi, Yaoqiang ; Tan, Qi ; Gong, Tao ; Li, Qing-yuan ; Zhu, Ya ; Duan, Xiaoqiong ; Yang, Chunhui ; Ding, Jia-wei ; Li, Shilin ; Xie, He ; Li, Yujia ; Chen, Limin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c326t-74163ffb835009d85af92115910def172d31777248720a587ff612d4224d670c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>Amplification</topic><topic>Analytical Chemistry</topic><topic>Antibodies</topic><topic>Characterization and Evaluation of Materials</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>CRISPR</topic><topic>Microengineering</topic><topic>Nanochemistry</topic><topic>Nanotechnology</topic><topic>Original Paper</topic><topic>Pathogens</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Shi, Yaoqiang</creatorcontrib><creatorcontrib>Tan, Qi</creatorcontrib><creatorcontrib>Gong, Tao</creatorcontrib><creatorcontrib>Li, Qing-yuan</creatorcontrib><creatorcontrib>Zhu, Ya</creatorcontrib><creatorcontrib>Duan, Xiaoqiong</creatorcontrib><creatorcontrib>Yang, Chunhui</creatorcontrib><creatorcontrib>Ding, Jia-wei</creatorcontrib><creatorcontrib>Li, Shilin</creatorcontrib><creatorcontrib>Xie, He</creatorcontrib><creatorcontrib>Li, Yujia</creatorcontrib><creatorcontrib>Chen, Limin</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>MEDLINE - Academic</collection><jtitle>Mikrochimica acta (1966)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Shi, Yaoqiang</au><au>Tan, Qi</au><au>Gong, Tao</au><au>Li, Qing-yuan</au><au>Zhu, Ya</au><au>Duan, Xiaoqiong</au><au>Yang, Chunhui</au><au>Ding, Jia-wei</au><au>Li, Shilin</au><au>Xie, He</au><au>Li, Yujia</au><au>Chen, Limin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cascaded signal amplification strategy for ultra-specific, ultra-sensitive, and visual detection of Shigella flexneri</atitle><jtitle>Mikrochimica acta (1966)</jtitle><stitle>Microchim Acta</stitle><addtitle>Mikrochim Acta</addtitle><date>2024-05-01</date><risdate>2024</risdate><volume>191</volume><issue>5</issue><spage>271</spage><epage>271</epage><pages>271-271</pages><artnum>271</artnum><issn>0026-3672</issn><eissn>1436-5073</eissn><abstract>Pathogen infections including
Shigella flexneri
have posed a significant threat to human health for numerous years. Although culturing and qPCR were the gold standards for pathogen detection, time-consuming and instrument-dependent restrict their application in rapid diagnosis and economically less-developed regions. Thus, it is urgently needed to develop rapid, simple, sensitive, accurate, and low-cost detection methods for pathogen detection. In this study, an immunomagnetic beads-recombinase polymerase amplification-CRISPR/Cas12a (IMB-RPA-CRISPR/Cas12a) method was built based on a cascaded signal amplification strategy for ultra-specific, ultra-sensitive, and visual detection of
S. flexneri
in the laboratory. Firstly,
S. flexneri
was specifically captured and enriched by IMB (
Shigella
antibody-coated magnetic beads), and the genomic DNA was released and used as the template in the RPA reaction. Then, the RPA products were mixed with the pre-loaded CRISPR/Cas12a for fluorescence visualization. The results were observed by naked eyes under LED blue light, with a sensitivity of 5 CFU/mL in a time of 70 min. With no specialized equipment or complicated technical requirements, the IMB-RPA-CRISPR/Cas12a diagnostic method can be used for visual, rapid, and simple detection of
S. flexneri
and can be easily adapted to monitoring other pathogens.
Graphical abstract</abstract><cop>Vienna</cop><pub>Springer Vienna</pub><pmid>38632191</pmid><doi>10.1007/s00604-024-06309-0</doi><tpages>1</tpages><orcidid>https://orcid.org/0000-0001-6076-7711</orcidid></addata></record> |
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| subjects | Amplification Analytical Chemistry Antibodies Characterization and Evaluation of Materials Chemistry Chemistry and Materials Science CRISPR Microengineering Nanochemistry Nanotechnology Original Paper Pathogens |
| title | Cascaded signal amplification strategy for ultra-specific, ultra-sensitive, and visual detection of Shigella flexneri |
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