A Rolling Circle-Amplified G‑Quadruplex/Hemin DNAzyme for Chemiluminescence Immunoassay of the SARS-CoV‑2 Protein
Sensitive detection of the SARS-CoV-2 protein remains a great research interest in clinical screening and diagnosis owing to the coronavirus epidemic. Here, an ultrasensitive chemiluminescence (CL) imaging strategy was developed through proximity hybridization to trigger the formation of a rolling c...
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Veröffentlicht in: | Analytical chemistry (Washington) 2021-07, Vol.93 (28), p.9933-9938 |
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description | Sensitive detection of the SARS-CoV-2 protein remains a great research interest in clinical screening and diagnosis owing to the coronavirus epidemic. Here, an ultrasensitive chemiluminescence (CL) imaging strategy was developed through proximity hybridization to trigger the formation of a rolling circle-amplified G-quadruplex/hemin DNAzyme for the detection of the SARS-CoV-2 protein. The target protein was first recognized by a pair of DNA–antibody conjugates, Ab-1 and Ab-2, to form a proximity-ligated complex, Ab-1/SARS-CoV-2/Ab-2, which contained a DNA sequence complemental to block DNA and thus induced a strand displacement reaction to release the primer from a block/primer complex. The released primer then triggered a rolling circle amplification to form abundant DNAzyme units in the presence of hemin, which produced a strong chemiluminescent signal for the detection of the target protein by catalyzing the oxidation of luminol by hydrogen peroxide. The proposed assay showed a detectable concentration range over 5 orders of magnitude with the detection limit down to 6.46 fg/mL. The excellent selectivity, simple procedure, acceptable accuracy, and intrinsic high throughput of the imaging technique for analysis of serum samples demonstrated the potential applicability of the proposed detection method in clinical screening and diagnosis. |
doi_str_mv | 10.1021/acs.analchem.1c02229 |
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Here, an ultrasensitive chemiluminescence (CL) imaging strategy was developed through proximity hybridization to trigger the formation of a rolling circle-amplified G-quadruplex/hemin DNAzyme for the detection of the SARS-CoV-2 protein. The target protein was first recognized by a pair of DNA–antibody conjugates, Ab-1 and Ab-2, to form a proximity-ligated complex, Ab-1/SARS-CoV-2/Ab-2, which contained a DNA sequence complemental to block DNA and thus induced a strand displacement reaction to release the primer from a block/primer complex. The released primer then triggered a rolling circle amplification to form abundant DNAzyme units in the presence of hemin, which produced a strong chemiluminescent signal for the detection of the target protein by catalyzing the oxidation of luminol by hydrogen peroxide. The proposed assay showed a detectable concentration range over 5 orders of magnitude with the detection limit down to 6.46 fg/mL. The excellent selectivity, simple procedure, acceptable accuracy, and intrinsic high throughput of the imaging technique for analysis of serum samples demonstrated the potential applicability of the proposed detection method in clinical screening and diagnosis.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.1c02229</identifier><identifier>PMID: 34227801</identifier><language>eng</language><publisher>Washington: American Chemical Society</publisher><subject>Amplification ; Antibodies ; Chemiluminescence ; Chemistry ; Coronaviruses ; COVID-19 ; Deoxyribonucleic acid ; Diagnosis ; DNA ; Hemin ; Hybridization ; Hydrogen peroxide ; Imaging techniques ; Immunoassay ; Nucleotide sequence ; Oxidation ; Proteins ; Screening ; Selectivity ; Severe acute respiratory syndrome coronavirus 2 ; Target detection ; Target recognition ; Viral diseases</subject><ispartof>Analytical chemistry (Washington), 2021-07, Vol.93 (28), p.9933-9938</ispartof><rights>2021 American Chemical Society</rights><rights>Copyright American Chemical Society Jul 20, 2021</rights><rights>2021 American Chemical Society 2021 American Chemical Society</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a454t-8927b38a9dcb88c164464a45fe9004078344b73e76d8d3ebe3a8e753e61fa8793</citedby><cites>FETCH-LOGICAL-a454t-8927b38a9dcb88c164464a45fe9004078344b73e76d8d3ebe3a8e753e61fa8793</cites><orcidid>0000-0002-6741-5302 ; 0000-0003-1379-122X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/acs.analchem.1c02229$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/acs.analchem.1c02229$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>230,314,780,784,885,2765,27076,27924,27925,56738,56788</link.rule.ids></links><search><creatorcontrib>Zhang, Rui</creatorcontrib><creatorcontrib>Wu, Jie</creatorcontrib><creatorcontrib>Ao, Hang</creatorcontrib><creatorcontrib>Fu, Jinling</creatorcontrib><creatorcontrib>Qiao, Bin</creatorcontrib><creatorcontrib>Wu, Qiang</creatorcontrib><creatorcontrib>Ju, Huangxian</creatorcontrib><title>A Rolling Circle-Amplified G‑Quadruplex/Hemin DNAzyme for Chemiluminescence Immunoassay of the SARS-CoV‑2 Protein</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>Sensitive detection of the SARS-CoV-2 protein remains a great research interest in clinical screening and diagnosis owing to the coronavirus epidemic. Here, an ultrasensitive chemiluminescence (CL) imaging strategy was developed through proximity hybridization to trigger the formation of a rolling circle-amplified G-quadruplex/hemin DNAzyme for the detection of the SARS-CoV-2 protein. The target protein was first recognized by a pair of DNA–antibody conjugates, Ab-1 and Ab-2, to form a proximity-ligated complex, Ab-1/SARS-CoV-2/Ab-2, which contained a DNA sequence complemental to block DNA and thus induced a strand displacement reaction to release the primer from a block/primer complex. The released primer then triggered a rolling circle amplification to form abundant DNAzyme units in the presence of hemin, which produced a strong chemiluminescent signal for the detection of the target protein by catalyzing the oxidation of luminol by hydrogen peroxide. The proposed assay showed a detectable concentration range over 5 orders of magnitude with the detection limit down to 6.46 fg/mL. The excellent selectivity, simple procedure, acceptable accuracy, and intrinsic high throughput of the imaging technique for analysis of serum samples demonstrated the potential applicability of the proposed detection method in clinical screening and diagnosis.</description><subject>Amplification</subject><subject>Antibodies</subject><subject>Chemiluminescence</subject><subject>Chemistry</subject><subject>Coronaviruses</subject><subject>COVID-19</subject><subject>Deoxyribonucleic acid</subject><subject>Diagnosis</subject><subject>DNA</subject><subject>Hemin</subject><subject>Hybridization</subject><subject>Hydrogen peroxide</subject><subject>Imaging techniques</subject><subject>Immunoassay</subject><subject>Nucleotide sequence</subject><subject>Oxidation</subject><subject>Proteins</subject><subject>Screening</subject><subject>Selectivity</subject><subject>Severe acute respiratory syndrome coronavirus 2</subject><subject>Target detection</subject><subject>Target recognition</subject><subject>Viral diseases</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><recordid>eNp9kctu1DAUhi0EosPAG7CwxIZNpseOEzsbpGiAtlLFpQW2lpOcdFw58dROEMOKV-AVeRI8mqESLFhZ8n_xOf4Iec5gxYCzU9PGlRmNazc4rFgLnPPqAVmwgkNWKsUfkgUA5BmXACfkSYy3AIwBKx-Tk1xwLhWwBZlreuWds-MNXdvQOszqYetsb7GjZ79-_Pw4my7MW4ffTs9xsCN9_a7-vhuQ9j7QdXraujldY2xxbJFeDMM8ehOj2VHf02mD9Lq-us7W_ksq4_RD8BPa8Sl51BsX8dnxXJLPb998Wp9nl-_PLtb1ZWZEIaZMVVw2uTJV1zZKtawUohRJ6rECECBVLkQjc5Rlp7ocG8yNQlnkWLLeKFnlS_Lq0LudmwG7NOMUjNPbYAcTdtobq_9WRrvRN_6rVlyWZfq9JXl5LAj-bsY46cGmVZ0zI_o5al4IVUEBuUrWF_9Yb_0cEqC9q5C8kEqUySUOrjb4GAP298Mw0HuuOnHVf7jqI9cUg0Nsr973_jfyG6HoqlI</recordid><startdate>20210720</startdate><enddate>20210720</enddate><creator>Zhang, Rui</creator><creator>Wu, Jie</creator><creator>Ao, Hang</creator><creator>Fu, Jinling</creator><creator>Qiao, Bin</creator><creator>Wu, Qiang</creator><creator>Ju, Huangxian</creator><general>American Chemical Society</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0002-6741-5302</orcidid><orcidid>https://orcid.org/0000-0003-1379-122X</orcidid></search><sort><creationdate>20210720</creationdate><title>A Rolling Circle-Amplified G‑Quadruplex/Hemin DNAzyme for Chemiluminescence Immunoassay of the SARS-CoV‑2 Protein</title><author>Zhang, Rui ; Wu, Jie ; Ao, Hang ; Fu, Jinling ; Qiao, Bin ; Wu, Qiang ; Ju, Huangxian</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a454t-8927b38a9dcb88c164464a45fe9004078344b73e76d8d3ebe3a8e753e61fa8793</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Amplification</topic><topic>Antibodies</topic><topic>Chemiluminescence</topic><topic>Chemistry</topic><topic>Coronaviruses</topic><topic>COVID-19</topic><topic>Deoxyribonucleic acid</topic><topic>Diagnosis</topic><topic>DNA</topic><topic>Hemin</topic><topic>Hybridization</topic><topic>Hydrogen peroxide</topic><topic>Imaging techniques</topic><topic>Immunoassay</topic><topic>Nucleotide sequence</topic><topic>Oxidation</topic><topic>Proteins</topic><topic>Screening</topic><topic>Selectivity</topic><topic>Severe acute respiratory syndrome coronavirus 2</topic><topic>Target detection</topic><topic>Target recognition</topic><topic>Viral diseases</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhang, Rui</creatorcontrib><creatorcontrib>Wu, Jie</creatorcontrib><creatorcontrib>Ao, Hang</creatorcontrib><creatorcontrib>Fu, Jinling</creatorcontrib><creatorcontrib>Qiao, Bin</creatorcontrib><creatorcontrib>Wu, Qiang</creatorcontrib><creatorcontrib>Ju, Huangxian</creatorcontrib><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhang, Rui</au><au>Wu, Jie</au><au>Ao, Hang</au><au>Fu, Jinling</au><au>Qiao, Bin</au><au>Wu, Qiang</au><au>Ju, Huangxian</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A Rolling Circle-Amplified G‑Quadruplex/Hemin DNAzyme for Chemiluminescence Immunoassay of the SARS-CoV‑2 Protein</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2021-07-20</date><risdate>2021</risdate><volume>93</volume><issue>28</issue><spage>9933</spage><epage>9938</epage><pages>9933-9938</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><abstract>Sensitive detection of the SARS-CoV-2 protein remains a great research interest in clinical screening and diagnosis owing to the coronavirus epidemic. Here, an ultrasensitive chemiluminescence (CL) imaging strategy was developed through proximity hybridization to trigger the formation of a rolling circle-amplified G-quadruplex/hemin DNAzyme for the detection of the SARS-CoV-2 protein. The target protein was first recognized by a pair of DNA–antibody conjugates, Ab-1 and Ab-2, to form a proximity-ligated complex, Ab-1/SARS-CoV-2/Ab-2, which contained a DNA sequence complemental to block DNA and thus induced a strand displacement reaction to release the primer from a block/primer complex. The released primer then triggered a rolling circle amplification to form abundant DNAzyme units in the presence of hemin, which produced a strong chemiluminescent signal for the detection of the target protein by catalyzing the oxidation of luminol by hydrogen peroxide. The proposed assay showed a detectable concentration range over 5 orders of magnitude with the detection limit down to 6.46 fg/mL. The excellent selectivity, simple procedure, acceptable accuracy, and intrinsic high throughput of the imaging technique for analysis of serum samples demonstrated the potential applicability of the proposed detection method in clinical screening and diagnosis.</abstract><cop>Washington</cop><pub>American Chemical Society</pub><pmid>34227801</pmid><doi>10.1021/acs.analchem.1c02229</doi><tpages>6</tpages><orcidid>https://orcid.org/0000-0002-6741-5302</orcidid><orcidid>https://orcid.org/0000-0003-1379-122X</orcidid><oa>free_for_read</oa></addata></record> |
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subjects | Amplification Antibodies Chemiluminescence Chemistry Coronaviruses COVID-19 Deoxyribonucleic acid Diagnosis DNA Hemin Hybridization Hydrogen peroxide Imaging techniques Immunoassay Nucleotide sequence Oxidation Proteins Screening Selectivity Severe acute respiratory syndrome coronavirus 2 Target detection Target recognition Viral diseases |
title | A Rolling Circle-Amplified G‑Quadruplex/Hemin DNAzyme for Chemiluminescence Immunoassay of the SARS-CoV‑2 Protein |
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