A sensitive and specific antigen detection assay for Middle East respiratory syndrome coronavirus
Since its emergence in 2012, more than 900 laboratory-confirmed cases of Middle East respiratory syndrome (MERS) have been reported with a fatality rate of more than 30%. However, no antigen detection assay for commercial use is available for diagnosis. In this study, the full-length nucleocapsid pr...
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| Veröffentlicht in: | Emerging microbes & infections 2015-04, Vol.4 (1), p.1-5 |
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| creator | Chen, Yixin Chan, Kwok-Hung Kang, Yahong Chen, Honglin Luk, Hayes KH Poon, Rosana WS Chan, Jasper FW Yuen, Kwok-Yung Xia, Ningshao Lau, Susanna KP Woo, Patrick CY |
| description | Since its emergence in 2012, more than 900 laboratory-confirmed cases of Middle East respiratory syndrome (MERS) have been reported with a fatality rate of more than 30%. However, no antigen detection assay for commercial use is available for diagnosis. In this study, the full-length nucleocapsid protein (NP) gene of MERS coronavirus (MERS-CoV) was cloned and expressed in Escherichia coli. A MERS-CoV NP capture enzyme-linked immunosorbent assay (ELISA) using two MERS-CoV-NP-specific monoclonal antibodies (MAbs) generated was developed. The ELISA was evaluated using 129 nasopharyngeal aspirates (NPAs) positive for various respiratory viruses and simulated positive NPAs by adding serial dilutions of MERS-CoV. Using a cutoff OD of 0.19, all 129 NPAs positive for respiratory viruses showed very low OD, with a specificity of 100%. For the two simulated MERS-CoV-positive NPAs with serial dilutions of live MERS-CoV, all samples with ≥10 50% tissue culture infective dose (TCID
50
)/0.1 mL showed positive results. For the 10 additional NPAs with 20 and 200 TCID
50
/0.1 mL of live MERS-CoV added, all were positive. A highly sensitive and specific MAbs-based antigen capture ELISA has been developed for MERS. This sensitive and specific antigen capture ELISA should be useful for detection of MERS-CoV in human and dromedaries and in field studies. |
| doi_str_mv | 10.1038/emi.2015.26 |
| format | Article |
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50
)/0.1 mL showed positive results. For the 10 additional NPAs with 20 and 200 TCID
50
/0.1 mL of live MERS-CoV added, all were positive. A highly sensitive and specific MAbs-based antigen capture ELISA has been developed for MERS. This sensitive and specific antigen capture ELISA should be useful for detection of MERS-CoV in human and dromedaries and in field studies.</description><identifier>ISSN: 2222-1751</identifier><identifier>EISSN: 2222-1751</identifier><identifier>DOI: 10.1038/emi.2015.26</identifier><identifier>PMID: 26421268</identifier><language>eng</language><publisher>United States: Taylor & Francis</publisher><subject>Animals ; Antibodies, Monoclonal - biosynthesis ; Antibodies, Monoclonal - immunology ; antigen ; Antigens ; Antigens, Viral - analysis ; Camelus ; Coronavirus Infections - diagnosis ; Coronavirus Infections - immunology ; Coronaviruses ; detection ; Disease Reservoirs ; ELISA ; Enzyme-Linked Immunosorbent Assay - methods ; Enzyme-Linked Immunosorbent Assay - standards ; Female ; Humans ; MERS coronavirus ; Mice ; Mice, Inbred BALB C ; Middle East respiratory syndrome ; Middle East Respiratory Syndrome Coronavirus - immunology ; Middle East Respiratory Syndrome Coronavirus - isolation & purification ; Nucleocapsid Proteins - analysis ; Nucleocapsid Proteins - genetics ; Nucleocapsid Proteins - metabolism ; Original ; Respiratory diseases ; Sensitivity and Specificity ; Zoonoses - diagnosis ; Zoonoses - immunology</subject><ispartof>Emerging microbes & infections, 2015-04, Vol.4 (1), p.1-5</ispartof><rights>2015 The Author(s) 2015</rights><rights>Copyright Nature Publishing Group Apr 2015</rights><rights>2015 The Author(s). This work is licensed under the Creative Commons Attribution – Non-Commercial – Share Alike License http://creativecommons.org/licenses/by-nc-sa/3.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>Copyright © 2015 Shanghai Shangyixun Cultural Communication Co., Ltd 2015 Shanghai Shangyixun Cultural Communication Co., Ltd</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c484t-e9d7072c48608306497d6dc939b8dd946d38f89b178cec8424d3f52fad5d0cfa3</citedby><cites>FETCH-LOGICAL-c484t-e9d7072c48608306497d6dc939b8dd946d38f89b178cec8424d3f52fad5d0cfa3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4575394/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4575394/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,27502,27924,27925,53791,53793,59143,59144</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26421268$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chen, Yixin</creatorcontrib><creatorcontrib>Chan, Kwok-Hung</creatorcontrib><creatorcontrib>Kang, Yahong</creatorcontrib><creatorcontrib>Chen, Honglin</creatorcontrib><creatorcontrib>Luk, Hayes KH</creatorcontrib><creatorcontrib>Poon, Rosana WS</creatorcontrib><creatorcontrib>Chan, Jasper FW</creatorcontrib><creatorcontrib>Yuen, Kwok-Yung</creatorcontrib><creatorcontrib>Xia, Ningshao</creatorcontrib><creatorcontrib>Lau, Susanna KP</creatorcontrib><creatorcontrib>Woo, Patrick CY</creatorcontrib><title>A sensitive and specific antigen detection assay for Middle East respiratory syndrome coronavirus</title><title>Emerging microbes & infections</title><addtitle>Emerg Microbes Infect</addtitle><description>Since its emergence in 2012, more than 900 laboratory-confirmed cases of Middle East respiratory syndrome (MERS) have been reported with a fatality rate of more than 30%. However, no antigen detection assay for commercial use is available for diagnosis. In this study, the full-length nucleocapsid protein (NP) gene of MERS coronavirus (MERS-CoV) was cloned and expressed in Escherichia coli. A MERS-CoV NP capture enzyme-linked immunosorbent assay (ELISA) using two MERS-CoV-NP-specific monoclonal antibodies (MAbs) generated was developed. The ELISA was evaluated using 129 nasopharyngeal aspirates (NPAs) positive for various respiratory viruses and simulated positive NPAs by adding serial dilutions of MERS-CoV. Using a cutoff OD of 0.19, all 129 NPAs positive for respiratory viruses showed very low OD, with a specificity of 100%. For the two simulated MERS-CoV-positive NPAs with serial dilutions of live MERS-CoV, all samples with ≥10 50% tissue culture infective dose (TCID
50
)/0.1 mL showed positive results. For the 10 additional NPAs with 20 and 200 TCID
50
/0.1 mL of live MERS-CoV added, all were positive. A highly sensitive and specific MAbs-based antigen capture ELISA has been developed for MERS. This sensitive and specific antigen capture ELISA should be useful for detection of MERS-CoV in human and dromedaries and in field studies.</description><subject>Animals</subject><subject>Antibodies, Monoclonal - biosynthesis</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>antigen</subject><subject>Antigens</subject><subject>Antigens, Viral - analysis</subject><subject>Camelus</subject><subject>Coronavirus Infections - diagnosis</subject><subject>Coronavirus Infections - immunology</subject><subject>Coronaviruses</subject><subject>detection</subject><subject>Disease Reservoirs</subject><subject>ELISA</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Enzyme-Linked Immunosorbent Assay - standards</subject><subject>Female</subject><subject>Humans</subject><subject>MERS coronavirus</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Middle East respiratory syndrome</subject><subject>Middle East Respiratory Syndrome Coronavirus - immunology</subject><subject>Middle East Respiratory Syndrome Coronavirus - isolation & purification</subject><subject>Nucleocapsid Proteins - analysis</subject><subject>Nucleocapsid Proteins - genetics</subject><subject>Nucleocapsid Proteins - metabolism</subject><subject>Original</subject><subject>Respiratory diseases</subject><subject>Sensitivity and Specificity</subject><subject>Zoonoses - diagnosis</subject><subject>Zoonoses - immunology</subject><issn>2222-1751</issn><issn>2222-1751</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>0YH</sourceid><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><recordid>eNp9kc9rFDEUxwex2FJ78i4BL0LZNb9mklyEUloVKr3oOWSTl5oyk6xJZmX-e7NuLVXEd0ke-fDhvXy77hXBa4KZfAdTWFNM-jUdnnUntNWKiJ48f3I_7s5KucetBB444S-6YzpwSuggTzpzgQrEEmrYATLRobIFG3ywranhDiJyUMHWkCIypZgF-ZTR5-DcCOjKlIoylG3Ipqa8oLJEl9MEyKacotmFPJeX3ZE3Y4Gzh_O0-3p99eXy4-rm9sOny4ubleWS1xUoJ7CgrRmwZG1QJdzgrGJqI51TfHBMeqk2REgLVnLKHfM99cb1Dltv2Gn3_uDdzpsJnIVYsxn1NofJ5EUnE_SfLzF803dpp3kveqZ4E7x9EOT0fYZS9RSKhXE0EdJcNBFEMkYl7hv65i_0Ps05tvU05YwLwtvv_o8iQhGFeyX31PmBsjmVksE_jkyw3mesW8Z6n7H-5Xz9dMtH9neiDRgOQIgtqcn8SHl0upplTNlnE20omv3L_BPvXbVo</recordid><startdate>20150401</startdate><enddate>20150401</enddate><creator>Chen, Yixin</creator><creator>Chan, Kwok-Hung</creator><creator>Kang, Yahong</creator><creator>Chen, Honglin</creator><creator>Luk, Hayes KH</creator><creator>Poon, Rosana WS</creator><creator>Chan, Jasper FW</creator><creator>Yuen, Kwok-Yung</creator><creator>Xia, Ningshao</creator><creator>Lau, Susanna KP</creator><creator>Woo, Patrick CY</creator><general>Taylor & Francis</general><general>Taylor & Francis Ltd</general><general>Nature Publishing Group</general><scope>0YH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>COVID</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20150401</creationdate><title>A sensitive and specific antigen detection assay for Middle East respiratory syndrome coronavirus</title><author>Chen, Yixin ; Chan, Kwok-Hung ; Kang, Yahong ; Chen, Honglin ; Luk, Hayes KH ; Poon, Rosana WS ; Chan, Jasper FW ; Yuen, Kwok-Yung ; Xia, Ningshao ; Lau, Susanna KP ; Woo, Patrick CY</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c484t-e9d7072c48608306497d6dc939b8dd946d38f89b178cec8424d3f52fad5d0cfa3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal - biosynthesis</topic><topic>Antibodies, Monoclonal - immunology</topic><topic>antigen</topic><topic>Antigens</topic><topic>Antigens, Viral - analysis</topic><topic>Camelus</topic><topic>Coronavirus Infections - diagnosis</topic><topic>Coronavirus Infections - immunology</topic><topic>Coronaviruses</topic><topic>detection</topic><topic>Disease Reservoirs</topic><topic>ELISA</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Enzyme-Linked Immunosorbent Assay - standards</topic><topic>Female</topic><topic>Humans</topic><topic>MERS coronavirus</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Middle East respiratory syndrome</topic><topic>Middle East Respiratory Syndrome Coronavirus - immunology</topic><topic>Middle East Respiratory Syndrome Coronavirus - isolation & purification</topic><topic>Nucleocapsid Proteins - analysis</topic><topic>Nucleocapsid Proteins - genetics</topic><topic>Nucleocapsid Proteins - metabolism</topic><topic>Original</topic><topic>Respiratory diseases</topic><topic>Sensitivity and Specificity</topic><topic>Zoonoses - diagnosis</topic><topic>Zoonoses - immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chen, Yixin</creatorcontrib><creatorcontrib>Chan, Kwok-Hung</creatorcontrib><creatorcontrib>Kang, Yahong</creatorcontrib><creatorcontrib>Chen, Honglin</creatorcontrib><creatorcontrib>Luk, Hayes KH</creatorcontrib><creatorcontrib>Poon, Rosana WS</creatorcontrib><creatorcontrib>Chan, Jasper FW</creatorcontrib><creatorcontrib>Yuen, Kwok-Yung</creatorcontrib><creatorcontrib>Xia, Ningshao</creatorcontrib><creatorcontrib>Lau, Susanna KP</creatorcontrib><creatorcontrib>Woo, Patrick CY</creatorcontrib><collection>Access via Taylor & Francis (Open Access Collection)</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Coronavirus Research Database</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Emerging microbes & infections</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chen, Yixin</au><au>Chan, Kwok-Hung</au><au>Kang, Yahong</au><au>Chen, Honglin</au><au>Luk, Hayes KH</au><au>Poon, Rosana WS</au><au>Chan, Jasper FW</au><au>Yuen, Kwok-Yung</au><au>Xia, Ningshao</au><au>Lau, Susanna KP</au><au>Woo, Patrick CY</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A sensitive and specific antigen detection assay for Middle East respiratory syndrome coronavirus</atitle><jtitle>Emerging microbes & infections</jtitle><addtitle>Emerg Microbes Infect</addtitle><date>2015-04-01</date><risdate>2015</risdate><volume>4</volume><issue>1</issue><spage>1</spage><epage>5</epage><pages>1-5</pages><issn>2222-1751</issn><eissn>2222-1751</eissn><abstract>Since its emergence in 2012, more than 900 laboratory-confirmed cases of Middle East respiratory syndrome (MERS) have been reported with a fatality rate of more than 30%. However, no antigen detection assay for commercial use is available for diagnosis. In this study, the full-length nucleocapsid protein (NP) gene of MERS coronavirus (MERS-CoV) was cloned and expressed in Escherichia coli. A MERS-CoV NP capture enzyme-linked immunosorbent assay (ELISA) using two MERS-CoV-NP-specific monoclonal antibodies (MAbs) generated was developed. The ELISA was evaluated using 129 nasopharyngeal aspirates (NPAs) positive for various respiratory viruses and simulated positive NPAs by adding serial dilutions of MERS-CoV. Using a cutoff OD of 0.19, all 129 NPAs positive for respiratory viruses showed very low OD, with a specificity of 100%. For the two simulated MERS-CoV-positive NPAs with serial dilutions of live MERS-CoV, all samples with ≥10 50% tissue culture infective dose (TCID
50
)/0.1 mL showed positive results. For the 10 additional NPAs with 20 and 200 TCID
50
/0.1 mL of live MERS-CoV added, all were positive. A highly sensitive and specific MAbs-based antigen capture ELISA has been developed for MERS. This sensitive and specific antigen capture ELISA should be useful for detection of MERS-CoV in human and dromedaries and in field studies.</abstract><cop>United States</cop><pub>Taylor & Francis</pub><pmid>26421268</pmid><doi>10.1038/emi.2015.26</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Antibodies, Monoclonal - biosynthesis Antibodies, Monoclonal - immunology antigen Antigens Antigens, Viral - analysis Camelus Coronavirus Infections - diagnosis Coronavirus Infections - immunology Coronaviruses detection Disease Reservoirs ELISA Enzyme-Linked Immunosorbent Assay - methods Enzyme-Linked Immunosorbent Assay - standards Female Humans MERS coronavirus Mice Mice, Inbred BALB C Middle East respiratory syndrome Middle East Respiratory Syndrome Coronavirus - immunology Middle East Respiratory Syndrome Coronavirus - isolation & purification Nucleocapsid Proteins - analysis Nucleocapsid Proteins - genetics Nucleocapsid Proteins - metabolism Original Respiratory diseases Sensitivity and Specificity Zoonoses - diagnosis Zoonoses - immunology |
| title | A sensitive and specific antigen detection assay for Middle East respiratory syndrome coronavirus |
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