Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2
SARS-CoV-2 has emerged as a previously unknown zoonotic coronavirus that spread worldwide causing a serious pandemic. While reliable nucleic acid–based diagnostic assays were rapidly available, only a limited number of validated serological assays were available in the early phase of the pandemic. H...
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| Veröffentlicht in: | European journal of clinical microbiology & infectious diseases 2021-04, Vol.40 (4), p.751-759 |
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| container_title | European journal of clinical microbiology & infectious diseases |
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| creator | Lapuente, Dennis Maier, Clara Irrgang, Pascal Hübner, Julian Peter, Antonia Sophia Hoffmann, Markus Ensser, Armin Ziegler, Katharina Winkler, Thomas H. Birkholz, Torsten Kremer, Andreas E. Steininger, Philipp Korn, Klaus Neipel, Frank Überla, Klaus Tenbusch, Matthias |
| description | SARS-CoV-2 has emerged as a previously unknown zoonotic coronavirus that spread worldwide causing a serious pandemic. While reliable nucleic acid–based diagnostic assays were rapidly available, only a limited number of validated serological assays were available in the early phase of the pandemic. Here, we evaluated a novel flow cytometric approach to assess spike-specific antibody responses.HEK 293T cells expressing SARS-CoV-2 spike protein in its natural confirmation on the surface were used to detect specific IgG and IgM antibody responses in patient sera by flow cytometry. A soluble angiotensin-converting-enzyme 2 (ACE-2) variant was developed as external standard to quantify spike-specific antibody responses on different assay platforms. Analyses of 201 pre-COVID-19 sera proved a high assay specificity in comparison to commercially available CLIA and ELISA systems, while also revealing the highest sensitivity in specimens from PCR-confirmed SARS-CoV-2-infected patients. The external standard allowed robust quantification of antibody responses among different assay platforms. In conclusion, our newly established flow cytometric assay allows sensitive and quantitative detection of SARS-CoV-2-specific antibodies, which can be easily adopted in different laboratories and does not rely on external supply of assay kits. The flow cytometric assay also provides a blueprint for rapid development of serological tests to other emerging viral infections |
| doi_str_mv | 10.1007/s10096-020-04072-7 |
| format | Article |
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While reliable nucleic acid–based diagnostic assays were rapidly available, only a limited number of validated serological assays were available in the early phase of the pandemic. Here, we evaluated a novel flow cytometric approach to assess spike-specific antibody responses.HEK 293T cells expressing SARS-CoV-2 spike protein in its natural confirmation on the surface were used to detect specific IgG and IgM antibody responses in patient sera by flow cytometry. A soluble angiotensin-converting-enzyme 2 (ACE-2) variant was developed as external standard to quantify spike-specific antibody responses on different assay platforms. Analyses of 201 pre-COVID-19 sera proved a high assay specificity in comparison to commercially available CLIA and ELISA systems, while also revealing the highest sensitivity in specimens from PCR-confirmed SARS-CoV-2-infected patients. The external standard allowed robust quantification of antibody responses among different assay platforms. In conclusion, our newly established flow cytometric assay allows sensitive and quantitative detection of SARS-CoV-2-specific antibodies, which can be easily adopted in different laboratories and does not rely on external supply of assay kits. The flow cytometric assay also provides a blueprint for rapid development of serological tests to other emerging viral infections</description><identifier>ISSN: 0934-9723</identifier><identifier>EISSN: 1435-4373</identifier><identifier>DOI: 10.1007/s10096-020-04072-7</identifier><identifier>PMID: 33078221</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Angiotensin ; Angiotensin-Converting Enzyme 2 ; Antibodies ; Antibodies, Viral - immunology ; Assaying ; Biomedical and Life Sciences ; Biomedicine ; Coronaviruses ; COVID-19 ; COVID-19 - diagnosis ; COVID-19 Serological Testing - methods ; Enzyme-Linked Immunosorbent Assay ; Flow cytometry ; Flow Cytometry - methods ; HEK293 Cells ; Humans ; Immunoglobulin G ; Immunoglobulin G - immunology ; Immunoglobulin M ; Immunoglobulin M - immunology ; Internal Medicine ; Medical Microbiology ; Nucleic acids ; Original ; Original Article ; Pandemics ; Peptidyl-dipeptidase A ; Platforms ; Reference Standards ; Reproducibility of Results ; SARS-CoV-2 - immunology ; Sensitivity and Specificity ; Serological tests ; Serology ; Severe acute respiratory syndrome coronavirus 2 ; Spike Glycoprotein, Coronavirus - immunology ; Spike protein ; Viral diseases</subject><ispartof>European journal of clinical microbiology & infectious diseases, 2021-04, Vol.40 (4), p.751-759</ispartof><rights>The Author(s) 2020</rights><rights>The Author(s) 2020. 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While reliable nucleic acid–based diagnostic assays were rapidly available, only a limited number of validated serological assays were available in the early phase of the pandemic. Here, we evaluated a novel flow cytometric approach to assess spike-specific antibody responses.HEK 293T cells expressing SARS-CoV-2 spike protein in its natural confirmation on the surface were used to detect specific IgG and IgM antibody responses in patient sera by flow cytometry. A soluble angiotensin-converting-enzyme 2 (ACE-2) variant was developed as external standard to quantify spike-specific antibody responses on different assay platforms. Analyses of 201 pre-COVID-19 sera proved a high assay specificity in comparison to commercially available CLIA and ELISA systems, while also revealing the highest sensitivity in specimens from PCR-confirmed SARS-CoV-2-infected patients. The external standard allowed robust quantification of antibody responses among different assay platforms. In conclusion, our newly established flow cytometric assay allows sensitive and quantitative detection of SARS-CoV-2-specific antibodies, which can be easily adopted in different laboratories and does not rely on external supply of assay kits. The flow cytometric assay also provides a blueprint for rapid development of serological tests to other emerging viral infections</description><subject>Angiotensin</subject><subject>Angiotensin-Converting Enzyme 2</subject><subject>Antibodies</subject><subject>Antibodies, Viral - immunology</subject><subject>Assaying</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Coronaviruses</subject><subject>COVID-19</subject><subject>COVID-19 - diagnosis</subject><subject>COVID-19 Serological Testing - methods</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Flow cytometry</subject><subject>Flow Cytometry - methods</subject><subject>HEK293 Cells</subject><subject>Humans</subject><subject>Immunoglobulin G</subject><subject>Immunoglobulin G - immunology</subject><subject>Immunoglobulin M</subject><subject>Immunoglobulin M - immunology</subject><subject>Internal Medicine</subject><subject>Medical Microbiology</subject><subject>Nucleic acids</subject><subject>Original</subject><subject>Original Article</subject><subject>Pandemics</subject><subject>Peptidyl-dipeptidase A</subject><subject>Platforms</subject><subject>Reference Standards</subject><subject>Reproducibility of Results</subject><subject>SARS-CoV-2 - immunology</subject><subject>Sensitivity and Specificity</subject><subject>Serological tests</subject><subject>Serology</subject><subject>Severe acute respiratory syndrome coronavirus 2</subject><subject>Spike Glycoprotein, Coronavirus - immunology</subject><subject>Spike protein</subject><subject>Viral diseases</subject><issn>0934-9723</issn><issn>1435-4373</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kT9vFDEQxS1ERC6BL0CBLNGkMdhje51tIkUnCEiRIuUPBY3l9Y6TjfbWF9tHdN8ekwtHSJFmPNL83huPHiHvBf8kODefc61twzhwxhU3wMwrMhNKaqakka_JjLdSsdaA3CV7Od_yKjo05g3ZlbJ2AGJGfp675dDThHkZp4w0jPGe-nWJCyxp8NTl7NY0xETLDdIeC_oyxInGQN1Uhi7266040xLpxfH5BZvHHwzekp3gxozvHt99cvX1y-X8Gzs9O_k-Pz5lXitemDJd24S26TQEjx4PpUTfa0RwIoiu06EH35pQGUDnemwlmlZ03KlGK1RynxxtfJerboG9x6kkN9plGhYurW10g_1_Mg039jr-skYbEFpWg4NHgxTvVpiLXQzZ4zi6CeMqW1AaNAfVQEU_PkNv4ypN9TxbEck1VLhSsKF8ijknDNvPCG7_RGc30dkanX2Izpoq-vD0jK3kb1YVkBsg19F0jenf7hdsfwODmaW-</recordid><startdate>20210401</startdate><enddate>20210401</enddate><creator>Lapuente, Dennis</creator><creator>Maier, Clara</creator><creator>Irrgang, Pascal</creator><creator>Hübner, Julian</creator><creator>Peter, Antonia Sophia</creator><creator>Hoffmann, Markus</creator><creator>Ensser, Armin</creator><creator>Ziegler, Katharina</creator><creator>Winkler, Thomas H.</creator><creator>Birkholz, Torsten</creator><creator>Kremer, Andreas E.</creator><creator>Steininger, Philipp</creator><creator>Korn, Klaus</creator><creator>Neipel, Frank</creator><creator>Überla, Klaus</creator><creator>Tenbusch, Matthias</creator><general>Springer Berlin Heidelberg</general><general>Springer Nature B.V</general><scope>C6C</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>7QL</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0003-3951-9056</orcidid></search><sort><creationdate>20210401</creationdate><title>Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2</title><author>Lapuente, Dennis ; Maier, Clara ; Irrgang, Pascal ; Hübner, Julian ; Peter, Antonia Sophia ; Hoffmann, Markus ; Ensser, Armin ; Ziegler, Katharina ; Winkler, Thomas H. ; Birkholz, Torsten ; Kremer, Andreas E. ; Steininger, Philipp ; Korn, Klaus ; Neipel, Frank ; Überla, Klaus ; Tenbusch, Matthias</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c540t-47b96f96b52fcece833ecd5ee2a1f1bb5fd2c97f6f92eaade93e791b0a4654e43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Angiotensin</topic><topic>Angiotensin-Converting Enzyme 2</topic><topic>Antibodies</topic><topic>Antibodies, Viral - 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In conclusion, our newly established flow cytometric assay allows sensitive and quantitative detection of SARS-CoV-2-specific antibodies, which can be easily adopted in different laboratories and does not rely on external supply of assay kits. The flow cytometric assay also provides a blueprint for rapid development of serological tests to other emerging viral infections</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>33078221</pmid><doi>10.1007/s10096-020-04072-7</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0003-3951-9056</orcidid><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 0934-9723 |
| ispartof | European journal of clinical microbiology & infectious diseases, 2021-04, Vol.40 (4), p.751-759 |
| issn | 0934-9723 1435-4373 |
| language | eng |
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| source | MEDLINE; SpringerNature Journals |
| subjects | Angiotensin Angiotensin-Converting Enzyme 2 Antibodies Antibodies, Viral - immunology Assaying Biomedical and Life Sciences Biomedicine Coronaviruses COVID-19 COVID-19 - diagnosis COVID-19 Serological Testing - methods Enzyme-Linked Immunosorbent Assay Flow cytometry Flow Cytometry - methods HEK293 Cells Humans Immunoglobulin G Immunoglobulin G - immunology Immunoglobulin M Immunoglobulin M - immunology Internal Medicine Medical Microbiology Nucleic acids Original Original Article Pandemics Peptidyl-dipeptidase A Platforms Reference Standards Reproducibility of Results SARS-CoV-2 - immunology Sensitivity and Specificity Serological tests Serology Severe acute respiratory syndrome coronavirus 2 Spike Glycoprotein, Coronavirus - immunology Spike protein Viral diseases |
| title | Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2 |
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