Recommendations for Fully Vaccinated People
COVID-19 Homepage
Interim Guidelines for COVID-19 Antibody Testing
Interim Guidelines for COVID-19 Antibody Testing in Clinical and Public Health Settings
Summary of Recent Changes
Key Points:
Antibody tests have public health value for monitoring and evaluating population levels of immunity, as well as clinical utility for patients.
- Antibody tests that have received Emergency Use Authorization (EUA) from the U.S. Food and Drug Administration (FDA) may be used for both public health and clinical purposes. Individual performance characteristics for each test can be found in the test’s instructions for use (IFU).
- Antibody testing should not be used to determine whether someone is currently infected with SARS-CoV-2. Viral tests detect current infection.
- Antibody testing is not currently recommended to assess for immunity to SARS-CoV-2 following COVID-19 vaccination or to assess the need for vaccination in an unvaccinated person.
When a person becomes infected with a pathogen, their immune system makes antibodies specifically to fight it. A person’s immune system can also safely learn to make antibodies through vaccination. Once you have antibodies to a particular disease, they provide some protection from that disease.
Even if a person does get sick, having antibodies can protect them from getting severely ill because their body has some experience in fighting that disease. How long this protection lasts can be different for each disease, each person, or influenced by other factors. Antibodies are just one part of a persons’ immune response.
Antibodies to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes COVID-19, can be detected in the blood of people who have recovered from COVID-19 or people who have been vaccinated against COVID-19. It is important to remember that some people with antibodies to SARS-CoV-2 may become infected after vaccination (vaccine breakthrough infection) or after recovering from a past infection (reinfection). For many diseases, including COVID-19, antibodies are expected to decrease or “wane” over time. As their antibodies wane, a person may become more vulnerable to SARS-CoV-2 infection. Even after a person’s antibodies wane, their immune system may have cells that remember the virus and that can act quickly to protect the person from severe illness if they become infected.
Antibody tests can detect the presence of these antibodies in serum within days to weeks following acute infection or vaccination. Antibody tests are not used for diagnosing a current case of COVID-19. Everyone, regardless of whether they have antibodies or not, should still take steps to protect themselves and others, including staying up to date on vaccination.
Antibody tests can be used to:
- Determine if a person has COVID-19 antibodies, which suggests past infection or vaccination.*
- Aid in the diagnosis of multisystem inflammatory syndrome in children (MIS-C) and in adults (MIS-A).
- Monitor and evaluate population levels of immunity.
*Antibody tests are not recommended or authorized by the FDA to assess someone’s immunity after COVID-19 vaccination or determine if they need to be vaccinated. However, EUA indications do not preclude use of antibody tests in vaccinated individuals in certain situations.
Antibody tests should not be used to:
- Diagnose current infection.**
- Determine if someone can return to work or school.
- Group people together in settings such as schools, dormitories, and correctional facilities; or to exempt someone from screening testing.
- Exempt a person who wears personal protective equipment (PPE) at work from following site-specific requirements
**Acute infection from SARS-CoV-2 is determined best by diagnostic testing using a nucleic acid amplification test (NAAT) or antigen test.
Choice of antibody test and testing strategy
For all clinical and public health purposes, it is recommended to use one of the numerous antibody tests for SARS-CoV-2 that have been authorized by FDA. The list of qualitative and semi-quantitative SARS-CoV-2 antibody tests granted an EUA by the FDA can be found on FDA’s website.
Antibody tests with very high sensitivity and specificity are preferred since they are more likely to exhibit high positive (probability that the person testing positive actually has antibodies) and negative predictive values (probability that the person testing negative actually does not have antibodies) when administered at least 3 weeks after the onset of illness.
Additional considerations when selecting an antibody test include:
- IgG levels appear to decrease more slowly over time than levels of other classes of antibody. Therefore, assays that measure total antibody or IgG could have higher sensitivity than IgM assays as more time passes since a person’s last infection.
- IgM antibody can persist for weeks to months following infection, though its persistence appears to be shorter than IgG’s; therefore, detection of IgM could suggest relatively recent infection.
- Detection of persistent antibodies varies by the test used.
FDA has issued an EUA for surrogate neutralization tests, which are qualitative binding assays that detect antibodies that block the interaction between the virus and the cellular virus receptor (ACE-2) without using cells or infectious virus. Plaque reduction neutralization assays are considered the gold standard for detection of neutralizing antibodies, but require cells, infectious virus, and are difficult to standardize. Although the surrogate neutralization test exhibits correlation to a plaque reduction neutralization test, the clinical or public health applicability has not been established.
The clinical applicability of semi-quantitative tests has not been established.
Interpretation of anti-S and anti-N antibody results based on vaccination status
Vaccination status
Vaccination status
Anti-S antibody
Anti-S antibody
Anti-N antibody
Anti-N antibody
Interpretation*
Interpretation*
Vaccinated
Vaccinated
+
+
+
+
Vaccinated and previously infected
Vaccinated and previously infected
Vaccinated
Vaccinated
+
+
–
–
Vaccinated and not previously infected
Vaccinated and not previously infected
Unvaccinated
Unvaccinated
+
+
+
+
Not vaccinated and previously infected
Not vaccinated and previously infected
Unvaccinated
Unvaccinated
–
–
–
–
Not previously vaccinated or infected
Not previously vaccinated or infected
Unknown
Unknown
+
+
+
+
Previously infected, may or may not have been vaccinated
Previously infected, may or may not have been vaccinated
Unknown
Unknown
+
+
–
–
Vaccinated with no previous infection
Vaccinated with no previous infection
Unknown
Unknown
–
–
–
–
Not previously vaccinated or infected
Not previously vaccinated or infected
*Potential false positive or false negative results, failure to develop detectable antibodies after vaccination or infection, and waning of antibodies with time after infection or vaccination should be considered when interpreting antibody test results.
Antibody tests can be used in seroprevalence studies to estimate vaccine coverage, or immunity from infection or vaccination in a community.
A negative antibody test does not rule out previous infection. A proportion of persons who are infected with SARS-CoV-2 might not develop measurable antibodies, thereby limiting the sensitivity of any antibody test to detect previous infection in these individuals. Also, the extent to which seroreversion occurs varies according to the antibody test used. In addition, measurable antibodies also can wane over time.
All eligible people should be vaccinated and stay up to date on vaccination, including unvaccinated people who have previously been infected and have detectable antibodies.
Indications for antibody testing
Persons suspected of having COVID-19 who test positive by direct viral detection methods for SARS-CoV-2 (e.g., NAAT or antigen detection tests) typically begin to develop measurable antibody 7–14 days after illness onset, and by 3 weeks most persons will test positive for antibody. During this interval, the sensitivity of detecting infection using NAAT or antigen detection testing decreases and the sensitivity of serologic testing increases. Antibody testing may be useful to support the diagnosis of COVID-19 illness or complications of COVID-19 in the following situations:
- A positive antibody test at least 7 days following acute illness onset in persons who had a previous negative antibody test (e.g., seroconversion) but did not receive a positive viral test might indicate SARS-CoV-2 infection between the dates of the negative and positive antibody tests.
- A positive antibody test can help support a diagnosis when patients present with complications of COVID-19, such as multisystem inflammatory syndrome or other post-acute sequelae of COVID-19.
Although current EUA indications do not preclude the use of these tests in vaccinated individuals, none of the currently authorized tests have been specifically authorized to assess immunity or protection of people who have received a COVID-19 vaccine, including people with immunocompromising conditions.
Whether the test has been validated to specifically detect antibodies against the antigens employed by the test and whether the antigens cross-react with antibodies to antigens that are not employed by the test should be considered. The results of available anti-SARS-CoV-2 IgG antibody tests may be interpreted in the following way:
In a person never vaccinated:
- Testing positive for antibody against N (nucleocapsid protein), S (spike protein), or RBD (receptor-binding domain of S protein) indicates prior infection.
In vaccinated people:
- Testing positive for antibody against the vaccine antigen target, such as the S protein, while testing negative for other antigens (e.g., N) suggests that they have produced vaccine-induced antibody. They may have never been infected with SARS-CoV-2, or they may have had a previous infection, but the N protein antibodies have since waned.
- Testing positive for antibodies other than the vaccine-induced antibody, such as the N protein, indicates resolving or past SARS-CoV-2 infection that could have occurred before or after vaccination.
- Antibody testing is currently not recommended to assess for immunity to SARS-CoV-2 following COVID-19 vaccination.
Current vaccines distributed in the United States induce antibodies to S protein. The presence of antibodies to N protein indicates previous infection regardless of a person’s vaccination status, while presence of antibodies to S protein indicates either previous infection or vaccination. Since vaccines induce antibodies to specific viral protein targets, post-vaccination antibody test results will be negative in persons without a history of previous infection if the test used does not detect antibodies induced by the vaccine.
Antibody Testing in the United States
Antigenic targets
The choice of antigenic targets might help address different aspects of immune response. Antibody detection against receptor-binding domain (RBD) is considered to have higher correlation with functional aspects like ability to neutralize virus (6). Differential reactivity of S and N specific antibodies might be used to help differentiate previous infection from vaccination in serologic studies, particularly for vaccines that produce antibodies only against S protein (1, 25, 40).
Multiple forms of S protein—full-length (S1+S2) or partial (S1 domain or RBD)—are used as antigens for antibody tests. S protein is essential for virus entry into cells and is present on the viral surface. Within the S protein, the RBD is more conserved than S1 or full-length S.
N protein is the most abundantly expressed immunodominant protein and is more conserved across coronaviruses than S.
Types of antibody testing
Different types of assays can be used to determine different aspects of the adaptive immune response and functionality of antibodies. The tests can be broadly classified to detect either binding or neutralizing antibodies.
Performance of antibody tests
Independently evaluated test performance and the approval status of tests are listed on an FDA website. Most authorized tests are qualitative (providing a result that is positive, negative, or indeterminate) or semi-quantitative (providing a numerical result using a scale that is unique to that assay and not comparable to other assays); however, authorized quantitative assays (providing a measured and scaled assessment of antibody levels) are also available.
FDA requires commercially marketed antibody tests for SARS-CoV-2 to receive Emergency Use Authorization (EUA) or approval. Multiple agencies—including FDA, the National Cancer Institute/National Institutes of Health (NCI/NIH), CDC, and the Biomedical Advanced Research and Development Authority (BARDA)—are collaborating with members of academia and the medical community to evaluate the performance of antibody tests independently using a well-characterized set of clinical specimens (serum and plasma) collected before and during the COVID-19 pandemic.
The World Health Organization has developed international standards for SARS-CoV-2 antibody tests that can serve as the foundation for the calibration of tests that quantify antibodies. Both laboratory and point-of-care antibody tests have received EUA from the FDA. Antibody testing technologies include single-use lateral flow tests where the presence of antibody is demonstrated by a color change on a paper strip (similar to a pregnancy test) and laboratory-based immunoassays that allow for processing of many specimens at the same time. The EUA letter of authorization includes the settings in which each test is authorized, based on FDA’s determination of appropriate settings for use during the public health emergency.
References
- Hall VJ, Foulkes S, Charlett A, Atti A, Monk EJM, Simmons R, et al. SARS-CoV-2 infection rates of antibody-positive compared with antibody-negative health-care workers in England: a large, multicentre, prospective cohort study (SIREN). Lancet. 2021 Apr 17;397(10283):1459-69.
- Lumley SF, O’Donnell D, Stoesser NE, Matthews PC, Howarth A, Hatch SB, et al. Antibody status and incidence of SARS-CoV-2 infection in health care workers. N Engl J Med. 2020 Dec 23;384:533-40.
- Abdool Karim SS, de Oliveira T. New SARS-CoV-2 Variants – Clinical, Public Health, and Vaccine Implications. N Engl J Med. 2021 May 13;384(19):1866-8.
- Grifoni A, Weiskopf D, Ramirez SI, Mateus J, Dan JM, Moderbacher CR, et al. Targets of T cell responses to SARS-CoV-2 coronavirus in humans with COVID-19 disease and unexposed individuals. Cell. 2020 Jun 25;181(7):1489-501 e15.
- Robbiani DF, Gaebler C, Muecksch F, Lorenzi JCC, Wang Z, Cho A, et al. Convergent antibody responses to SARS-CoV-2 in convalescent individuals. Nature. 2020 Aug;584(7821):437-42.
- Suthar MS, Zimmerman MG, Kauffman RC, Mantus G, Linderman SL, Hudson WH, et al. Rapid generation of neutralizing antibody responses in COVID-19 patients. Cell Rep Med. 2020 Jun 23;1(3):100040.
- Qu J, Wu C, Li X, Zhang G, Jiang Z, Li X, et al. Profile of immunoglobulin G and IgM antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Clin Infect Dis. 2020 Nov 19;71(16):2255-8.
- Wolfel R, Corman VM, Guggemos W, Seilmaier M, Zange S, Muller MA, et al. Virological assessment of hospitalized patients with COVID-2019. Nature. 2020 May;581(7809):465-9.
- Iyer AS, Jones FK, Nodoushani A, Kelly M, Becker M, Slater D, et al. Persistence and decay of human antibody responses to the receptor binding domain of SARS-CoV-2 spike protein in COVID-19 patients. Sci Immunol. 2020 Oct 8;5(52).
- Matuchansky C. Mucosal immunity to SARS-CoV-2: a clinically relevant key to deciphering natural and vaccine-induced defences. Clin Microbiol Infect. 2021 Aug 12.
- Dan JM, Mateus J, Kato Y, Hastie KM, Faliti CE, Ramirez SI, et al. Immunological memory to SARS-CoV-2 assessed for greater than six months after infection. bioRxiv. 2020(10.1101/2020.11.15.383323).
- Milani GP, Dioni L, Favero C, Cantone L, Macchi C, Delbue S, et al. Serological follow-up of SARS-CoV-2 asymptomatic subjects. Sci Rep. 2020 Nov 18;10(1):20048.
- Rijkers G, Murk JL, Wintermans B, van Looy B, van den Berge M, Veenemans J, et al. Differences in antibody kinetics and functionality between severe and mild severe acute respiratory syndrome coronavirus 2 infections. J Infect Dis. 2020 Sep 14;222(8):1265-9.
- Choe PG, Kang CK, Suh HJ, Jung J, Kang E, Lee SY, et al. Antibody responses to SARS-CoV-2 at 8 weeks postinfection in asymptomatic patients. Emerg Infect Dis. 2020 Jun 24;26(10):2484-7.
- Petersen LR, Sami S, Vuong N, Pathela P, Weiss D, Morgenthau BM, et al. Lack of antibodies to SARS-CoV-2 in a large cohort of previously infected persons. Clin Infect Dis. 2020 Nov 4.
- Kaufman HW, Chen Z, Meyer WA, 3rd, Wohlgemuth JG. Insights from patterns of SARS-CoV-2 immunoglobulin G serology test results from a national clinical laboratory, United States, March-July 2020. Popul Health Manag. 2020 Nov 19.
- Payne DC, Smith-Jeffcoat SE, Nowak G, Chukwuma U, Geibe JR, Hawkins RJ, et al. SARS-CoV-2 Infections and serologic responses from a sample of U.S. Navy service members – USS Theodore Roosevelt, April 2020. MMWR Morb Mortal Wkly Rep. 2020 Jun 12;69(23):714-21.
- Ogega CO, Skinner NE, Blair PW, Park HS, Littlefield K, Ganesan A, et al. Durable SARS-CoV-2 B cell immunity after mild or severe disease. J Clin Invest. 2021 Apr 1;131(7).
- Gundlapalli AV, Reynolds MS, Brooks JT, Francisco A, Petersen L, McDonald LC, et al. SARS-CoV-2 serologic assay needs for the next phase of U.S. COVID-19 pandemic response. Open Forum Infecti Dis. 2020 17 Nov 2020;8:ofaa555.
- Selhorst P, Van Ierssel S, Michiels J, Marien J, Bartholomeeusen K, Dirinck E, et al. Symptomatic SARS-CoV-2 reinfection of a health care worker in a Belgian nosocomial outbreak despite primary neutralizing antibody response. Clin Infect Dis. 2020 Dec 14.
- To KK, Tsang OT, Leung WS, Tam AR, Wu TC, Lung DC, et al. Temporal profiles of viral load in posterior oropharyngeal saliva samples and serum antibody responses during infection by SARS-CoV-2: an observational cohort study. Lancet Infect Dis. 2020 May;20(5):565-74.
- Addetia A, Crawford KHD, Dingens A, Zhu H, Roychoudhury P, Huang ML, et al. Neutralizing antibodies correlate with protection from SARS-CoV-2 in humans during a fishery vessel outbreak with a high attack rate. J Clin Microbiol. 2020 Oct 21;58(11).
- Pray IW, Gibbons-Burgener SN, Rosenberg AZ, Cole D, Borenstein S, Bateman A, et al. COVID-19 outbreak at an overnight summer school retreat – Wisconsin, July-August 2020. MMWR Morb Mortal Wkly Rep. 2020 Oct 30;69(43):1600-4.
- Jeffery-Smith A, Iyanger N, Williams SV, Chow JY, Aiano F, Hoschler K, et al. Antibodies to SARS-CoV-2 protect against re-infection during outbreaks in care homes, September and October 2020. Euro Surveill. 2021 Feb;26(5).
- Harvey RA, Rassen JA, Kabelac CA, Turenne W, Leonard S, Klesh R, et al. Association of SARS-CoV-2 Seropositive Antibody Test With Risk of Future Infection. JAMA Intern Med. 2021 May 1;181(5):672-9.
- Letizia AG, Ge Y, Goforth CW, Weir DL, Lizewski R, Lizewski S, et al. SARS-CoV-2 seropositivity among US Marine recruits attending basic training, United States, Spring-Fall 2020. Emerg Infect Dis. 2021 Feb 2;27(4).
- Chandrashekar A, Liu J, Martinot AJ, McMahan K, Mercado NB, Peter L, et al. SARS-CoV-2 infection protects against rechallenge in rhesus macaques. Science. 2020 Aug 14;369(6505):812-7.
- Feng, S. Phillips, D. White, T., et al. Correlates of protection against symptomatic and asymptomatic SARS-CoV-2 infection. medRxiv 2021. .
- Gilbert PB, Montefiori DC, McDermott A, Fong Y, Benkeser DC, Deng W, et al. Immune Correlates Analysis of the mRNA-1273 COVID-19 Vaccine Efficacy Trial. medRxiv. 2021 Aug 15.
- Edridge AWD, Kaczorowska J, Hoste ACR, Bakker M, Klein M, Loens K, et al. Seasonal coronavirus protective immunity is short-lasting. Nat Med. 2020 Nov;26(11):1691-3.
- Alshukairi AN, Khalid I, Ahmed WA, Dada AM, Bayumi DT, Malic LS, et al. Antibody response and disease severity in healthcare worker MERS survivors. Emerg Infect Dis. 2016 Jun;22(6).
- Wu LP, Wang NC, Chang YH, Tian XY, Na DY, Zhang LY, et al. Duration of antibody responses after severe acute respiratory syndrome. Emerg Infect Dis. 2007 Oct;13(10):1562-4.
- Payne DC, Iblan I, Rha B, Alqasrawi S, Haddadin A, Al Nsour M, et al. Persistence of antibodies against Middle East respiratory syndrome coronavirus. Emerg Infect Dis. 2016 Oct;22(10):1824-6.
- Cromer D, Juno JA, Khoury D, Reynaldi A, Wheatley AK, Kent SJ, et al. Prospects for durable immune control of SARS-CoV-2 and prevention of reinfection. Nat Rev Immunol. 2021 Jun;21(6):395-404.
- Poland GA, Ovsyannikova IG, Kennedy RB. SARS-CoV-2 immunity: review and applications to phase 3 vaccine candidates. Lancet. 2020 Nov 14;396(10262):1595-606.
- Baden LR, El Sahly HM, Essink B, Kotloff K, Frey S, Novak R, et al. Efficacy and Safety of the mRNA-1273 SARS-CoV-2 Vaccine. N Engl J Med. 2020 Dec 30.
- Polack FP, Thomas SJ, Kitchin N, Absalon J, Gurtman A, Lockhart S, et al. Safety and efficacy of the BNT162b2 mRNA COVID-19 vaccine. N Engl J Med. 2020 Dec 10.
- Thompson MG, Burgess JL, Naleway AL, Tyner HL, Yoon SK, Meece J, et al. Interim Estimates of Vaccine Effectiveness of BNT162b2 and mRNA-1273 COVID-19 Vaccines in Preventing SARS-CoV-2 Infection Among Health Care Personnel, First Responders, and Other Essential and Frontline Workers – Eight U.S. Locations, December 2020-March 2021. MMWR Morb Mortal Wkly Rep. 2021 Apr 2;70(13):495-500.
- Krammer F, Srivastava K, Alshammary H, Amoako AA, Awawda MH, Beach KF, et al. Antibody Responses in Seropositive Persons after a Single Dose of SARS-CoV-2 mRNA Vaccine. N Engl J Med. 2021 Apr 8;384(14):1372-4.
- Fotis C, Meimetis N, Tsolakos N, Politou M, Akinosoglou K, Pliaka V, et al. Accurate SARS-CoV-2 seroprevalence surveys require robust multi-antigen assays. Sci Rep. 2021 Mar 23;11(1):6614.
- Letizia AG, Ge Y, Vangeti S, Goforth C, Weir DL, Kuzmina NA, et al. SARS-CoV-2 seropositivity and subsequent infection risk in healthy young adults: a prospective cohort study. medRxiv. 2021.
- McMahan K, Yu J, Mercado NB, Loos C, Tostanoski LH, Chandrashekar A, et al. Correlates of protection against SARS-CoV-2 in rhesus macaques. Nature. 2021 Feb;590(7847):630-4.
- Cohen MS, Nirula A, Mulligan MJ, Novak RM, Marovich M, Yen C, et al. Effect of Bamlanivimab vs Placebo on Incidence of COVID-19 Among Residents and Staff of Skilled Nursing and Assisted Living Facilities: A Randomized Clinical Trial. JAMA. 2021 Jun 3.
- Regeneron. Regeneron reports positive interim data with regen-cov™ antibody cocktail used as passive vaccine to prevent COVID-19. Accessed June 4, 2021. Available at: https://investor.regeneron.com/news-releases/news-release-details/regeneron-reports-positive-interim-data-regen-covtm-antibody.
- Stephens DS, McElrath MJ. COVID-19 and the Path to Immunity. JAMA. 2020 Oct 6;324(13):1279-81.
- Public Health England. SARS-CoV-2 variants of concern and variants under investigation in England. Technical briefing 19. Accessed: Aug 20 2021. Available at: https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/1005517/Technical_Briefing_19.pdf.