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Performance of nucleocapsid and spike-based SARS-CoV-2 serologic assays.
PLoS One. 2020; 15(11):e0237828.Plos

Abstract

There is an urgent need for an accurate antibody test for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We have developed 3 ELISA methods, trimer spike IgA, trimer spike IgG, and nucleocapsid IgG, for detecting anti-SARS-CoV-2 antibodies. We evaluated their performance along with four commercial ELISAs, EDI™ Novel Coronavirus COVID-19 ELISA IgG and IgM, Euroimmun Anti-SARS-CoV-2 ELISA IgG and IgA, and one lateral flow assay, DPP® COVID-19 IgM/IgG System (Chembio). Both sensitivity and specificity were evaluated and the probable causes of false-positive reactions were determined. The assays were evaluated using 300 pre-epidemic samples and 100 PCR-confirmed COVID-19 samples. The sensitivities and specificities of the assays were as follows: 90%/100% (in-house trimer spike IgA), 90%/99.3% (in-house trimer spike IgG), 89%/98.3% (in-house nucleocapsid IgG), 73.7%/100% (EDI nucleocapsid IgM), 84.5%/95.1% (EDI nucleocapsid IgG), 95%/93.7% (Euroimmun S1 IgA), 82.8%/99.7% (Euroimmun S1 IgG), 82.0%/91.7% (Chembio nucleocapsid IgM), 92%/93.3% (Chembio nucleocapsid IgG). The presumed causes of false positive results from pre-epidemic samples in commercial and in-house assays were mixed. In some cases, assays lacked reproducibility. In other cases, reactivity was abrogated by competitive inhibition (spiking the sample with the same antigen that was used for coating ELISAs prior to performing the assay), suggesting positive reaction could be attributed to the presence of antibodies against these antigens. In other cases, reactivity was consistently detected but not abrogated by the spiking, suggesting positive reaction was not attributed to the presence of antibodies against these antigens. Overall, there was wide variability in assay performance using our samples, with in-house tests exhibiting the highest combined sensitivity and specificity. The causes of "false positivity" in pre-epidemic samples may be due to plasma antibodies apparently reacting with the corresponding antigen, or spurious reactivity may be directed against non-specific components in the assay system. Identification of these targets will be essential to improving assay performance.

Authors+Show Affiliations

Division of Clinical Care and Research, Institute of Human Virology, University of Maryland, Baltimore, Maryland, United States of America. Division of Vaccine Research, Institute of Human Virology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America.Division of Clinical Care and Research, Institute of Human Virology, University of Maryland, Baltimore, Maryland, United States of America. Division of Vaccine Research, Institute of Human Virology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America.Division of Epidemiology & Prevention, Institute of Human Virology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America.Division of Clinical Care and Research, Institute of Human Virology, University of Maryland, Baltimore, Maryland, United States of America.Division of Epidemiology & Prevention, Institute of Human Virology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America.Division of Vaccine Research, Institute of Human Virology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America.Department of Epidemiology and Public Health, University of Maryland School of Medicine Baltimore, Maryland, United States of America.Division of Vaccine Research, Institute of Human Virology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America.Division of Clinical Care and Research, Institute of Human Virology, University of Maryland, Baltimore, Maryland, United States of America.Division of Clinical Care and Research, Institute of Human Virology, University of Maryland, Baltimore, Maryland, United States of America. Division of Vaccine Research, Institute of Human Virology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America. Department of Medicine, Baltimore VA Medical Center, Baltimore, Maryland, United States of America.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

33137138

Citation

Rikhtegaran Tehrani, Zahra, et al. "Performance of Nucleocapsid and Spike-based SARS-CoV-2 Serologic Assays." PloS One, vol. 15, no. 11, 2020, pp. e0237828.
Rikhtegaran Tehrani Z, Saadat S, Saleh E, et al. Performance of nucleocapsid and spike-based SARS-CoV-2 serologic assays. PLoS One. 2020;15(11):e0237828.
Rikhtegaran Tehrani, Z., Saadat, S., Saleh, E., Ouyang, X., Constantine, N., DeVico, A. L., Harris, A. D., Lewis, G. K., Kottilil, S., & Sajadi, M. M. (2020). Performance of nucleocapsid and spike-based SARS-CoV-2 serologic assays. PloS One, 15(11), e0237828. https://doi.org/10.1371/journal.pone.0237828
Rikhtegaran Tehrani Z, et al. Performance of Nucleocapsid and Spike-based SARS-CoV-2 Serologic Assays. PLoS One. 2020;15(11):e0237828. PubMed PMID: 33137138.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Performance of nucleocapsid and spike-based SARS-CoV-2 serologic assays. AU - Rikhtegaran Tehrani,Zahra, AU - Saadat,Saman, AU - Saleh,Ebtehal, AU - Ouyang,Xin, AU - Constantine,Niel, AU - DeVico,Anthony L, AU - Harris,Anthony D, AU - Lewis,George K, AU - Kottilil,Shyam, AU - Sajadi,Mohammad M, Y1 - 2020/11/02/ PY - 2020/08/06/received PY - 2020/10/15/accepted PY - 2020/11/2/entrez PY - 2020/11/3/pubmed PY - 2020/11/20/medline SP - e0237828 EP - e0237828 JF - PloS one JO - PLoS One VL - 15 IS - 11 N2 - There is an urgent need for an accurate antibody test for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We have developed 3 ELISA methods, trimer spike IgA, trimer spike IgG, and nucleocapsid IgG, for detecting anti-SARS-CoV-2 antibodies. We evaluated their performance along with four commercial ELISAs, EDI™ Novel Coronavirus COVID-19 ELISA IgG and IgM, Euroimmun Anti-SARS-CoV-2 ELISA IgG and IgA, and one lateral flow assay, DPP® COVID-19 IgM/IgG System (Chembio). Both sensitivity and specificity were evaluated and the probable causes of false-positive reactions were determined. The assays were evaluated using 300 pre-epidemic samples and 100 PCR-confirmed COVID-19 samples. The sensitivities and specificities of the assays were as follows: 90%/100% (in-house trimer spike IgA), 90%/99.3% (in-house trimer spike IgG), 89%/98.3% (in-house nucleocapsid IgG), 73.7%/100% (EDI nucleocapsid IgM), 84.5%/95.1% (EDI nucleocapsid IgG), 95%/93.7% (Euroimmun S1 IgA), 82.8%/99.7% (Euroimmun S1 IgG), 82.0%/91.7% (Chembio nucleocapsid IgM), 92%/93.3% (Chembio nucleocapsid IgG). The presumed causes of false positive results from pre-epidemic samples in commercial and in-house assays were mixed. In some cases, assays lacked reproducibility. In other cases, reactivity was abrogated by competitive inhibition (spiking the sample with the same antigen that was used for coating ELISAs prior to performing the assay), suggesting positive reaction could be attributed to the presence of antibodies against these antigens. In other cases, reactivity was consistently detected but not abrogated by the spiking, suggesting positive reaction was not attributed to the presence of antibodies against these antigens. Overall, there was wide variability in assay performance using our samples, with in-house tests exhibiting the highest combined sensitivity and specificity. The causes of "false positivity" in pre-epidemic samples may be due to plasma antibodies apparently reacting with the corresponding antigen, or spurious reactivity may be directed against non-specific components in the assay system. Identification of these targets will be essential to improving assay performance. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/33137138/Performance_of_nucleocapsid_and_spike_based_SARS_CoV_2_serologic_assays_ L2 - https://dx.plos.org/10.1371/journal.pone.0237828 DB - PRIME DP - Unbound Medicine ER -