Ultrasensitive assay for saliva-based SARS-CoV-2 antigen detection
Widespread SARS-CoV-2 testing is invaluable for identifying asymptomatic/pre-symptomatic individuals. There remains a technological gap for highly reliable, easy, and quick SARS-CoV-2 diagnostic tests suitable for frequent mass testing. Compared to nasopharyngeal (NP) swab-based tests, saliva-based...
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| Veröffentlicht in: | Clinical chemistry and laboratory medicine 2022-04, Vol.60 (5), p.771-777 |
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| creator | Ren, Annie Sohaei, Dorsa Ulndreaj, Antigona Pons-Belda, Oscar D. Fernandez-Uriarte, Amaia Zacharioudakis, Ioannis Sigal, George B. Stengelin, Martin Mathew, Anu Campbell, Christopher Padmanabhan, Nikhil Romero, Daniel Joe, Jessica Soosaipillai, Antoninus Kulasingam, Vathany Mazzulli, Tony Li, Xinliu A. McGeer, Allison Diamandis, Eleftherios P. Prassas, Ioannis |
| description | Widespread SARS-CoV-2 testing is invaluable for identifying asymptomatic/pre-symptomatic individuals. There remains a technological gap for highly reliable, easy, and quick SARS-CoV-2 diagnostic tests suitable for frequent mass testing. Compared to nasopharyngeal (NP) swab-based tests, saliva-based methods are attractive due to easier and safer sampling. Current saliva-based SARS-CoV-2 rapid antigen tests (RATs) are hindered by limited analytical sensitivity. Here, we report one of the first ultrasensitive, saliva-based SARS-CoV-2 antigen assays with an analytical sensitivity of |
| doi_str_mv | 10.1515/cclm-2021-1142 |
| format | Article |
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Using the novel electrochemiluminescence (ECL)-based immunoassay, we measured the SARS-CoV-2 nucleocapsid (N) antigen concentration in 105 salivas, obtained from non-COVID-19 and COVID-19 patients. We then verified the results with a second, independent cohort of 689 patients (3.8% SARS-CoV-2 positivity rate). We also compared our method with a widely used point-of-care rapid test.
In the first cohort, at 100% specificity, the sensitivity was 92%. Our assay correctly identified samples with viral loads up to 35 CT cycles by saliva-based PCR. Paired NP swab-based PCR results were obtained for 86 cases. Our assay showed high concordance with saliva-based and NP swab-based PCR in samples with negative (<0.32 pg/mL) and strongly positive (>2 pg/mL) N antigen concentrations. In the second cohort, at 100% specificity, sensitivity was also 92%. Our assay is about 700-fold more sensitive than the Abbott Panbio Rapid Test.
We demonstrated the ultrasensitivity and specificity assay and its concordance with PCR. This novel assay is especially valuable when compliance to frequent swabbing may be problematic.</description><identifier>ISSN: 1434-6621</identifier><identifier>EISSN: 1437-4331</identifier><identifier>DOI: 10.1515/cclm-2021-1142</identifier><identifier>PMID: 35170269</identifier><language>eng</language><publisher>Germany: De Gruyter</publisher><subject>Antigens ; Antigens, Viral ; Coronaviruses ; COVID-19 ; COVID-19 - diagnosis ; COVID-19 Testing ; Electrochemiluminescence ; electrochemiluminescence (ECL) immunoassay ; Humans ; Immunoassay ; mass testing ; nasopharyngeal (NP) swab ; Nasopharynx ; Nucleocapsids ; Saliva ; Saliva - chemistry ; SARS-CoV-2 ; sensitivity ; Sensitivity analysis ; Sensitivity and Specificity ; Severe acute respiratory syndrome ; Severe acute respiratory syndrome coronavirus 2</subject><ispartof>Clinical chemistry and laboratory medicine, 2022-04, Vol.60 (5), p.771-777</ispartof><rights>2022 Walter de Gruyter GmbH, Berlin/Boston.</rights><rights>2022 Walter de Gruyter GmbH, Berlin/Boston</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c416t-543af8c7861cdb079583311ef19cd82b5917fa49d2fbc20e821581b4868109393</citedby><cites>FETCH-LOGICAL-c416t-543af8c7861cdb079583311ef19cd82b5917fa49d2fbc20e821581b4868109393</cites><orcidid>0000-0002-1589-820X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.degruyter.com/document/doi/10.1515/cclm-2021-1142/pdf$$EPDF$$P50$$Gwalterdegruyter$$H</linktopdf><linktohtml>$$Uhttps://www.degruyter.com/document/doi/10.1515/cclm-2021-1142/html$$EHTML$$P50$$Gwalterdegruyter$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,66497,68281</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35170269$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ren, Annie</creatorcontrib><creatorcontrib>Sohaei, Dorsa</creatorcontrib><creatorcontrib>Ulndreaj, Antigona</creatorcontrib><creatorcontrib>Pons-Belda, Oscar D.</creatorcontrib><creatorcontrib>Fernandez-Uriarte, Amaia</creatorcontrib><creatorcontrib>Zacharioudakis, Ioannis</creatorcontrib><creatorcontrib>Sigal, George B.</creatorcontrib><creatorcontrib>Stengelin, Martin</creatorcontrib><creatorcontrib>Mathew, Anu</creatorcontrib><creatorcontrib>Campbell, Christopher</creatorcontrib><creatorcontrib>Padmanabhan, Nikhil</creatorcontrib><creatorcontrib>Romero, Daniel</creatorcontrib><creatorcontrib>Joe, Jessica</creatorcontrib><creatorcontrib>Soosaipillai, Antoninus</creatorcontrib><creatorcontrib>Kulasingam, Vathany</creatorcontrib><creatorcontrib>Mazzulli, Tony</creatorcontrib><creatorcontrib>Li, Xinliu A.</creatorcontrib><creatorcontrib>McGeer, Allison</creatorcontrib><creatorcontrib>Diamandis, Eleftherios P.</creatorcontrib><creatorcontrib>Prassas, Ioannis</creatorcontrib><title>Ultrasensitive assay for saliva-based SARS-CoV-2 antigen detection</title><title>Clinical chemistry and laboratory medicine</title><addtitle>Clin Chem Lab Med</addtitle><description>Widespread SARS-CoV-2 testing is invaluable for identifying asymptomatic/pre-symptomatic individuals. There remains a technological gap for highly reliable, easy, and quick SARS-CoV-2 diagnostic tests suitable for frequent mass testing. Compared to nasopharyngeal (NP) swab-based tests, saliva-based methods are attractive due to easier and safer sampling. Current saliva-based SARS-CoV-2 rapid antigen tests (RATs) are hindered by limited analytical sensitivity. Here, we report one of the first ultrasensitive, saliva-based SARS-CoV-2 antigen assays with an analytical sensitivity of <0.32 pg/mL, corresponding to four viral RNA copies/µL, which is comparable to that of PCR-based tests.
Using the novel electrochemiluminescence (ECL)-based immunoassay, we measured the SARS-CoV-2 nucleocapsid (N) antigen concentration in 105 salivas, obtained from non-COVID-19 and COVID-19 patients. We then verified the results with a second, independent cohort of 689 patients (3.8% SARS-CoV-2 positivity rate). We also compared our method with a widely used point-of-care rapid test.
In the first cohort, at 100% specificity, the sensitivity was 92%. Our assay correctly identified samples with viral loads up to 35 CT cycles by saliva-based PCR. Paired NP swab-based PCR results were obtained for 86 cases. Our assay showed high concordance with saliva-based and NP swab-based PCR in samples with negative (<0.32 pg/mL) and strongly positive (>2 pg/mL) N antigen concentrations. In the second cohort, at 100% specificity, sensitivity was also 92%. Our assay is about 700-fold more sensitive than the Abbott Panbio Rapid Test.
We demonstrated the ultrasensitivity and specificity assay and its concordance with PCR. This novel assay is especially valuable when compliance to frequent swabbing may be problematic.</description><subject>Antigens</subject><subject>Antigens, Viral</subject><subject>Coronaviruses</subject><subject>COVID-19</subject><subject>COVID-19 - diagnosis</subject><subject>COVID-19 Testing</subject><subject>Electrochemiluminescence</subject><subject>electrochemiluminescence (ECL) immunoassay</subject><subject>Humans</subject><subject>Immunoassay</subject><subject>mass testing</subject><subject>nasopharyngeal (NP) swab</subject><subject>Nasopharynx</subject><subject>Nucleocapsids</subject><subject>Saliva</subject><subject>Saliva - chemistry</subject><subject>SARS-CoV-2</subject><subject>sensitivity</subject><subject>Sensitivity analysis</subject><subject>Sensitivity and Specificity</subject><subject>Severe acute respiratory syndrome</subject><subject>Severe acute respiratory syndrome coronavirus 2</subject><issn>1434-6621</issn><issn>1437-4331</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptkE1Lw0AQhhdRbK1ePUrAi5etO_uVXfCixS8QBGu9hs1mU1LSpO4mlf57E1sVxNMMzDPvDA9Cp0DGIEBcWlsuMSUUMACne2gInMWYMwb7Xz3HUlIYoKMQFoSAEDw-RAMmICZU6iG6mZWNN8FVoWiKtYtMCGYT5bWPgimLtcFpN8yi6fXLFE_qN0wjUzXF3FVR5hpnm6KujtFBbsrgTnZ1hGZ3t6-TB_z0fP84uX7CloNssODM5MrGSoLNUhJrobovweWgbaZoKjTEueE6o3lqKXGKglCQciUVEM00G6GLbe7K1--tC02yLIJ1ZWkqV7choZJqppSU0KHnf9BF3fqq-66juCSactIHjreU9XUI3uXJyhdL4zcJkKS3m_R2k95u0tvtFs52sW26dNkP_q2zA662wIcpG-czN_ftpmt-z_-fLImIY2CfAnqGig</recordid><startdate>20220426</startdate><enddate>20220426</enddate><creator>Ren, Annie</creator><creator>Sohaei, Dorsa</creator><creator>Ulndreaj, Antigona</creator><creator>Pons-Belda, Oscar D.</creator><creator>Fernandez-Uriarte, Amaia</creator><creator>Zacharioudakis, Ioannis</creator><creator>Sigal, George B.</creator><creator>Stengelin, Martin</creator><creator>Mathew, Anu</creator><creator>Campbell, Christopher</creator><creator>Padmanabhan, Nikhil</creator><creator>Romero, Daniel</creator><creator>Joe, Jessica</creator><creator>Soosaipillai, Antoninus</creator><creator>Kulasingam, Vathany</creator><creator>Mazzulli, Tony</creator><creator>Li, Xinliu A.</creator><creator>McGeer, Allison</creator><creator>Diamandis, Eleftherios P.</creator><creator>Prassas, Ioannis</creator><general>De Gruyter</general><general>Walter De Gruyter & Company</general><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>7QO</scope><scope>7T7</scope><scope>7TK</scope><scope>7U7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-1589-820X</orcidid></search><sort><creationdate>20220426</creationdate><title>Ultrasensitive assay for saliva-based SARS-CoV-2 antigen detection</title><author>Ren, Annie ; Sohaei, Dorsa ; Ulndreaj, Antigona ; Pons-Belda, Oscar D. ; Fernandez-Uriarte, Amaia ; Zacharioudakis, Ioannis ; Sigal, George B. ; Stengelin, Martin ; Mathew, Anu ; Campbell, Christopher ; Padmanabhan, Nikhil ; Romero, Daniel ; Joe, Jessica ; Soosaipillai, Antoninus ; Kulasingam, Vathany ; Mazzulli, Tony ; Li, Xinliu A. ; McGeer, Allison ; Diamandis, Eleftherios P. ; Prassas, Ioannis</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c416t-543af8c7861cdb079583311ef19cd82b5917fa49d2fbc20e821581b4868109393</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Antigens</topic><topic>Antigens, Viral</topic><topic>Coronaviruses</topic><topic>COVID-19</topic><topic>COVID-19 - diagnosis</topic><topic>COVID-19 Testing</topic><topic>Electrochemiluminescence</topic><topic>electrochemiluminescence (ECL) immunoassay</topic><topic>Humans</topic><topic>Immunoassay</topic><topic>mass testing</topic><topic>nasopharyngeal (NP) swab</topic><topic>Nasopharynx</topic><topic>Nucleocapsids</topic><topic>Saliva</topic><topic>Saliva - chemistry</topic><topic>SARS-CoV-2</topic><topic>sensitivity</topic><topic>Sensitivity analysis</topic><topic>Sensitivity and Specificity</topic><topic>Severe acute respiratory syndrome</topic><topic>Severe acute respiratory syndrome coronavirus 2</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ren, Annie</creatorcontrib><creatorcontrib>Sohaei, Dorsa</creatorcontrib><creatorcontrib>Ulndreaj, Antigona</creatorcontrib><creatorcontrib>Pons-Belda, Oscar D.</creatorcontrib><creatorcontrib>Fernandez-Uriarte, Amaia</creatorcontrib><creatorcontrib>Zacharioudakis, Ioannis</creatorcontrib><creatorcontrib>Sigal, George B.</creatorcontrib><creatorcontrib>Stengelin, Martin</creatorcontrib><creatorcontrib>Mathew, Anu</creatorcontrib><creatorcontrib>Campbell, Christopher</creatorcontrib><creatorcontrib>Padmanabhan, Nikhil</creatorcontrib><creatorcontrib>Romero, Daniel</creatorcontrib><creatorcontrib>Joe, Jessica</creatorcontrib><creatorcontrib>Soosaipillai, Antoninus</creatorcontrib><creatorcontrib>Kulasingam, Vathany</creatorcontrib><creatorcontrib>Mazzulli, Tony</creatorcontrib><creatorcontrib>Li, Xinliu A.</creatorcontrib><creatorcontrib>McGeer, Allison</creatorcontrib><creatorcontrib>Diamandis, Eleftherios P.</creatorcontrib><creatorcontrib>Prassas, Ioannis</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Neurosciences Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Clinical chemistry and laboratory medicine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ren, Annie</au><au>Sohaei, Dorsa</au><au>Ulndreaj, Antigona</au><au>Pons-Belda, Oscar D.</au><au>Fernandez-Uriarte, Amaia</au><au>Zacharioudakis, Ioannis</au><au>Sigal, George B.</au><au>Stengelin, Martin</au><au>Mathew, Anu</au><au>Campbell, Christopher</au><au>Padmanabhan, Nikhil</au><au>Romero, Daniel</au><au>Joe, Jessica</au><au>Soosaipillai, Antoninus</au><au>Kulasingam, Vathany</au><au>Mazzulli, Tony</au><au>Li, Xinliu A.</au><au>McGeer, Allison</au><au>Diamandis, Eleftherios P.</au><au>Prassas, Ioannis</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Ultrasensitive assay for saliva-based SARS-CoV-2 antigen detection</atitle><jtitle>Clinical chemistry and laboratory medicine</jtitle><addtitle>Clin Chem Lab Med</addtitle><date>2022-04-26</date><risdate>2022</risdate><volume>60</volume><issue>5</issue><spage>771</spage><epage>777</epage><pages>771-777</pages><issn>1434-6621</issn><eissn>1437-4331</eissn><abstract>Widespread SARS-CoV-2 testing is invaluable for identifying asymptomatic/pre-symptomatic individuals. There remains a technological gap for highly reliable, easy, and quick SARS-CoV-2 diagnostic tests suitable for frequent mass testing. Compared to nasopharyngeal (NP) swab-based tests, saliva-based methods are attractive due to easier and safer sampling. Current saliva-based SARS-CoV-2 rapid antigen tests (RATs) are hindered by limited analytical sensitivity. Here, we report one of the first ultrasensitive, saliva-based SARS-CoV-2 antigen assays with an analytical sensitivity of <0.32 pg/mL, corresponding to four viral RNA copies/µL, which is comparable to that of PCR-based tests.
Using the novel electrochemiluminescence (ECL)-based immunoassay, we measured the SARS-CoV-2 nucleocapsid (N) antigen concentration in 105 salivas, obtained from non-COVID-19 and COVID-19 patients. We then verified the results with a second, independent cohort of 689 patients (3.8% SARS-CoV-2 positivity rate). We also compared our method with a widely used point-of-care rapid test.
In the first cohort, at 100% specificity, the sensitivity was 92%. Our assay correctly identified samples with viral loads up to 35 CT cycles by saliva-based PCR. Paired NP swab-based PCR results were obtained for 86 cases. Our assay showed high concordance with saliva-based and NP swab-based PCR in samples with negative (<0.32 pg/mL) and strongly positive (>2 pg/mL) N antigen concentrations. In the second cohort, at 100% specificity, sensitivity was also 92%. Our assay is about 700-fold more sensitive than the Abbott Panbio Rapid Test.
We demonstrated the ultrasensitivity and specificity assay and its concordance with PCR. This novel assay is especially valuable when compliance to frequent swabbing may be problematic.</abstract><cop>Germany</cop><pub>De Gruyter</pub><pmid>35170269</pmid><doi>10.1515/cclm-2021-1142</doi><tpages>07</tpages><orcidid>https://orcid.org/0000-0002-1589-820X</orcidid><oa>free_for_read</oa></addata></record> |
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| ispartof | Clinical chemistry and laboratory medicine, 2022-04, Vol.60 (5), p.771-777 |
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| source | MEDLINE; De Gruyter journals |
| subjects | Antigens Antigens, Viral Coronaviruses COVID-19 COVID-19 - diagnosis COVID-19 Testing Electrochemiluminescence electrochemiluminescence (ECL) immunoassay Humans Immunoassay mass testing nasopharyngeal (NP) swab Nasopharynx Nucleocapsids Saliva Saliva - chemistry SARS-CoV-2 sensitivity Sensitivity analysis Sensitivity and Specificity Severe acute respiratory syndrome Severe acute respiratory syndrome coronavirus 2 |
| title | Ultrasensitive assay for saliva-based SARS-CoV-2 antigen detection |
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