Serology for SARS-CoV-2: Apprehensions, opportunities, and the path forward

Abstract

Serological testing for SARS-CoV-2 has enormous potential to contribute to COVID-19 pandemic response efforts. However, the required performance characteristics of antibody tests will critically depend on the use case (individual-level vs. population-level).

Use cases for SARS-CoV-2 serology

At the individual level, serologic tests are frequently used to support clinical diagnosis by determining recent or prior infection (to supplement PCR detection), or to determine vaccination status and requirements for boosting. In vaccine trials, individual assessments of antibody endpoints may be used to determine serostatus prior to enrollment, as a tool to reduce bias, simplify analyses, and minimize required sample sizes. (2) Specific to SARS-CoV-2, a widely discussed idea in the media has been the issuance of “immune passports” – the proposed use of serology to infer immunity and thus enable a person to work on the front lines or return to daily work routines. Such an application must be predicated on an established surrogate of protection – a given antibody endpoint associated with clinical protection from infection – and a test with sufficient specificity to ensure people are not unintentionally put in harm’s way. (3) Serology tests with relatively high but imperfect specificity may lead to substantial false-positive results when used in low incidence settings (Fig. 1A). For example, in a setting where 5% of the population has been infected, a test with 96% specificity and 90% sensitivity would lead to just 54% of positive results indicating a true infection (i.e., positive predictive value). In addition to the risks of false positives, false negatives may occur in some previously-infected persons who fail to produce antibodies specific to the antigens/epitopes in a given assay, or whose antibodies have already quickly waned, or, when used during an on-going epidemic, among those who have not yet mounted a specific antibody response. (4, 5) For these individuals that don’t mount a measurable antibody response despite having been infected, obtaining permission to return to work could be onerous. Further considerations that may undermine the individual use-case is that even with a true positive antibody result we do not know how well that translates to protection or immunity, nor whether those positive by an antibody test could still shed virus and infect others.

At the population level, representative cross-sectional serosurveys can provide aggregate ‘snapshots’ of infection history and immunity of a population. Understanding the proportion of the population infected by SARS-CoV-2 cannot be assessed based on PCR confirmed cases alone, due to variations in testing practices, timing of sampling, and the clinical spectrum of disease (e.g., asymptomatic infections). In contrast to case data, seroepidemiological datasets provide a less biased picture of risk of death (infection fatality rate), the amplitude of transmission in different populations, and can highlight disparities in infection rates without typical health-seeking behavior biases. Understanding age-specific or spatial distribution of susceptibility could guide policymakers about where to intervene and to what degree, by helping to answer questions such as: What IgG seroprevalence in children is acceptable to allow schools to open? Do infection attack rates differ between children and adults? Population-level surveys could also help estimate the probability and timing of future waves of disease, which will critically depend upon duration of immunity, (6) measure the impact of interventions such as physical distancing or vaccination, and in later stages, confirm the absence of transmission.

Here we underscore key differences between individual- and population-level use cases and emphasize that different use cases will require tests with different performance characteristics: while assays that “certify” an individual’s immunity need to be correlated with protection and have near-perfect specificity (to limit the number of false positives, when seroprevalence is low), assays to ascertain population-level exposure would have utility as long as the sensitivity and specificity are well-defined for the target population allowing for adjustment of seroprevalence estimates (Fig. 1B). Optimal thresholds for sensitivity/specificity can be “tuned” depending on local prevalence and intended use of the assay. For example, when conducting a serosurvey in low-prevalence settings, to achieve better precision of point estimates of disease burden, the assay specificity should be prioritized, typically at the cost of sensitivity. This can be achieved by raising the cutoff value for the assay used, for example by setting higher optical density readings as the threshold for positivity in an ELISA. Similarly, in a high-prevalence setting, test sensitivity should be prioritized at the cost of specificity. Thus, we recommend the consideration of multiple threshold values (cutoffs) for assays that can be flexibly used in different contexts.

Foundational studies to enhance the utility and interpretation of SARS-CoV-2 serology.

While many SARS-CoV-2 serological assays may have insufficient performance characteristics (sensitivity/specificity) to warrant use at the individual level, and the WHO currently recommends restricting antibody testing to research use only, we argue that these imperfect tests may nevertheless provide highly valuable tools to address critical public health questions, such as the safety of relaxing stay-at-home orders or school closures, or evaluations of alternative intervention measures. To fully realize the benefits of population-level seroepidemiological studies, a number of fundamental questions must be addressed, relating to test performance, the dynamics of antibody responses in relation to infection, and the link between antibody responses and immunity (Table 1). Answering these questions across different populations and epidemiologic contexts will require various study designs, which we view as key for optimal interpretation of the growing number of population-based, cross-sectional serological surveys.

Governance

Serosurveillance for SARS-CoV-2 will only be capable of contributing to actionable public health information if serology measurements flow into efficient data pipelines. Scale-up of serological testing for pandemic response must therefore be accompanied by a governance model at the sub-national, national and international levels, and by an operational research agenda that evaluates the utility of assays within specific contexts. With the plethora of new tests in development and diverse testing strategies, there is an urgent need for national-level strategies to enable pooling of results generated from different methods and sources. National-level governance will be required to provide oversight for sample collection and processing, linkage to personal data, and to coordinate results analysis to the appropriate scale for policy relevance. Much like a national census is translated into infrastructure appropriations, serosurveillance could be used for resource allocations (and future vaccination efforts) to target transmission hotspots.

Data from carefully designed serostudies are urgently needed prior to widespread adoption or implementation of antibody testing programs. To ensure comparison across studies, there is a need for harmonization of assay protocols, sharing of reference standards, and a set of best-practices for reporting results. Because seroepidemiological studies will require measurement of healthy individuals, various strategies for opportunistic sampling of individuals in community settings should be explored, as described in a proposed Global Serum Bank. (7) A host of ethical and privacy issues will need to be addressed; we suggest that serosurveillance platforms should incorporate broad consent, enabling future screening of serum collections for multiple biomarkers of public health concern beyond SARS-CoV-2 alone. The SARS-CoV-2 pandemic has highlighted the value of transparency in disease surveillance for all nations. We see a role for international coordination of national seroepidemiology programs to facilitate standardizing methods and dissemination of results among national public health laboratories.

In summary, seroepidemiological studies and integrated serosurveillance platforms are urgently needed to guide and tailor SARS-CoV-2 response efforts, and will continue to be critical for mitigating post-pandemic resurgence. Coordinated serosurveillance provides opportunities to combine control efforts for different diseases into one coordinated program; this may be particularly valuable to assess impact of the COVID-19 crisis on routine immunization programs. (8) Platforms should be designed with a longer term vision beyond COVID-19, to generate capacity for ‘precision public health’ to monitor additional major diseases, and provide insights into how disease occurrence is interrelated with other health risk factors. Finally, we stress that investing now in a fundamental and operational research agenda will allow us to rapidly develop serosurveillance as a powerful tool for population-level public health; however, the complexity of using serological assays within low prevalence settings to inform individual-based risk assessments – i.e., to inform decisions regarding return to work – is dangerously premature.

References and Notes

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   Wu F, Wang A, Liu M, et al. Neutralizing antibody responses to SARS-CoV-2 in a COVID-19 recovered patient cohort and their implications. medRxiv 2020; posted April 6, 2020. DOI: https://doi.org/. 10.1101/2020.03.30.20047365

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   Tan W, Lu Y, Zhang J, et al. Viral Kinetics and Antibody Responses in Patients with COVID-19. medRxiv 2020; posted April 20, 2020. DOI: https://doi.org/. 10.1101/2020.03.24.20042382

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