Evaluation of SARS-CoV-2 serology assays reveals a range of test performance

• Published in: Nature biotechnology 2020-10, Vol.38 (10), p.1174-1183
• Main Authors: Whitman, Jeffrey D., Hiatt, Joseph, Mowery, Cody T., Shy, Brian R., Yu, Ruby, Yamamoto, Tori N., Rathore, Ujjwal, Goldgof, Gregory M., Whitty, Caroline, Woo, Jonathan M., Gallman, Antonia E., Miller, Tyler E., Levine, Andrew G., Nguyen, David N., Bapat, Sagar P., Balcerek, Joanna, Bylsma, Sophia A., Lyons, Ana M., Li, Stacy, Wong, Allison Wai-yi, Gillis-Buck, Eva Mae, Steinhart, Zachary B., Lee, Youjin, Apathy, Ryan, Lipke, Mitchell J., Smith, Jennifer Anne, Zheng, Tina, Boothby, Ian C., Isaza, Erin, Chan, Jackie, Acenas, Dante D., Lee, Jinwoo, Macrae, Trisha A., Kyaw, Than S., Wu, David, Ng, Dianna L., Gu, Wei, York, Vanessa A., Eskandarian, Haig Alexander, Callaway, Perri C., Warrier, Lakshmi, Moreno, Mary E., Levan, Justine, Torres, Leonel, Farrington, Lila A., Loudermilk, Rita P., Koshal, Kanishka, Zorn, Kelsey C., Garcia-Beltran, Wilfredo F., Yang, Diane, Astudillo, Michael G., Bernstein, Bradley E., Gelfand, Jeffrey A., Ryan, Edward T., Charles, Richelle C., Iafrate, A. John, Lennerz, Jochen K., Miller, Steve, Chiu, Charles Y., Stramer, Susan L., Wilson, Michael R., Manglik, Aashish, Ye, Chun Jimmie, Krogan, Nevan J., Anderson, Mark S., Cyster, Jason G., Ernst, Joel D., Wu, Alan H. B., Lynch, Kara L., Bern, Caryn, Hsu, Patrick D., Marson, Alexander
• Format: Article
• Language: English
• Subjects: 692/699/255/2514; 692/700/139/1420; Adult; Aged; Aged, 80 and over; Agriculture; Antibodies; Antibodies, Viral - blood; Assaying; Betacoronavirus - genetics; Betacoronavirus - immunology; Betacoronavirus - isolation & purification; Biochemical assays; Bioinformatics; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; Chromatography, Affinity; Clinical Laboratory Techniques - methods; Clinical Laboratory Techniques - statistics & numerical data; Coronaviridae; Coronavirus Infections - diagnosis; Coronavirus Infections - epidemiology; Coronavirus Infections - immunology; Coronaviruses; COVID-19; COVID-19 Testing; Diagnosis; Enzyme-Linked Immunosorbent Assay; Enzymes; Evaluation; Female; Health aspects; Humans; Immunoassays; Immunoglobulin G; Immunoglobulin G - blood; Immunoglobulin M; Immunoglobulin M - blood; Life Sciences; Male; Middle Aged; Pandemics; Pneumonia, Viral - diagnosis; Pneumonia, Viral - epidemiology; Pneumonia, Viral - immunology; Point-of-Care Testing; Reverse Transcriptase Polymerase Chain Reaction; SARS-CoV-2; Sensitivity and Specificity; Serodiagnosis; Serological tests; Serology; Severe acute respiratory syndrome; Severe acute respiratory syndrome coronavirus 2; Type 2 diabetes; Viral diseases; Young Adult
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/s41587-020-0659-0

Appropriate use and interpretation of serological tests for assessments of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) exposure, infection and potential immunity require accurate data on assay performance. We conducted a head-to-head evaluation of ten point-of-care-style lateral flow assays (LFAs) and two laboratory-based enzyme-linked immunosorbent assays to detect anti-SARS-CoV-2 IgM and IgG antibodies in 5-d time intervals from symptom onset and studied the specificity of each assay in pre-coronavirus disease 2019 specimens. The percent of seropositive individuals increased with time, peaking in the latest time interval tested (>20 d after symptom onset). Test specificity ranged from 84.3% to 100.0% and was predominantly affected by variability in IgM results. LFA specificity could be increased by considering weak bands as negative, but this decreased detection of antibodies (sensitivity) in a subset of SARS-CoV-2 real-time PCR-positive cases. Our results underline the importance of seropositivity threshold determination and reader training for reliable LFA deployment. Although there was no standout serological assay, four tests achieved more than 80% positivity at later time points tested and more than 95% specificity. Of 12 serology assays tested, four detect antibodies in more than 80% of patients with COVID-19.