Evaluation of Serological SARS-CoV-2 Lateral Flow Assays for Rapid Point of Care Testing

Conklin, Steven E.; Martin, Kathryn; Manabe, Yukari C.; Schmidt, H; Keruly, Morgan; Klock, Ethan; Kirby, Charles; Baker, Owen R.; Fernández, Reinaldo E.; Eby, Yolanda; Hardick, Justin; Shaw‐Saliba, Kathryn; Rothman, Richard E.; Caturegli, Patrizio; Redd, Andrew R.; Tobian, Aaron A.R.; Bloch, Evan M.; Larman, H. Benjamin; Quinn, Thomas C.; Clarke, William; Laeyendecker, Oliver

doi:10.1101/2020.07.31.20166041

Cited by 21 publications

(22 citation statements)

References 25 publications

(28 reference statements)

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“…Finally, we characterized cross-reactive T cell responses in just 1 unexposed HD, because HD9 was the only unexposed HD in our cohort with preexisting immunity to SARS-CoV-2. Although we screened this individual using 2 different SARS-CoV-2 antibody tests with stated sensitivities of 100% among hospitalized patients by 3 weeks after symptom onset (24,25), negative findings do not definitively prove that this individual did not have asymptomatic infection. This is important, given the studies showing that seronegative exposed individuals can have SARS-specific T cell responses (7,26), although the cross-reactivity we describe here may also explain those results.…”

Section: Discussionmentioning

confidence: 98%

Healthy donor T cell responses to common cold coronaviruses and SARS-CoV-2

Woldemeskel

Kwaa

Garliss

et al. 2020

Journal of Clinical Investigation

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BACKGROUND. T cell responses to the common cold coronaviruses have not been well characterized. Preexisting T cell immunity to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been reported, and a recent study suggested that this immunity was due to cross-recognition of the novel coronavirus by T cells specific for the common cold coronaviruses. METHODS. We used the enzyme-linked immunospot (ELISPOT) assay to characterize the T cell responses against peptide pools derived from the spike protein of 3 common cold coronaviruses (HCoV-229E, HCoV-NL63, and HCoV-OC43) and SARS-CoV-2 in 21 healthy donors (HDs) who were seronegative for SARS-CoV-2 and had no known exposure to the virus. An in vitro expansion culture assay was also used to analyze memory T cell responses. RESULTS. We found responses to the spike protein of the 3 common cold coronaviruses in many of the donors. We then focused on HCoV-NL63 and detected broad T cell responses to the spike protein and identified 22 targeted peptides. Interestingly, only 1 study participant had a significant response to SARS-CoV-2 spike or nucleocapsid protein in the ELISPOT assay. In vitro expansion studies suggested that T cells specific for the HCoV-NL63 spike protein in this individual could also recognize SARS-CoV-2 spike protein peptide pools. CONCLUSION. HDs have circulating T cells specific for the spike proteins of HCoV-NL63, HCoV-229E, and HCoV-OC43. T cell responses to SARS-CoV-2 spike and nucleocapsid proteins were present in only 1 participant and were potentially the result of cross-recognition by T cells specific for the common cold coronaviruses. Further studies are needed to determine whether this cross-recognition influences coronavirus disease 2019 (COVID-19) outcomes.

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Section: Discussionmentioning

confidence: 98%

Healthy donor T cell responses to common cold coronaviruses and SARS-CoV-2

Woldemeskel

Kwaa

Garliss

et al. 2020

Journal of Clinical Investigation

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“…Drawbacks of LFAs include their higher cost-per-test rate, their inability to analyze multiple samples simultaneously, their general lack of quantitative data, and importantly, a several-fold reduced sensitivity when compared to non-rapid testing methods (71,72). Although RDTs are theoretically ideal for POC usage, recent studies have demonstrated that many newly developed RDTs for SARS-CoV-2 have failed to meet the necessary standards for sensitivity and specificity when compared to non-rapid testing (71)(72)(73)(74)(75)(76). Therefore, for research purposes, LFAs are not the ideal choice.…”

Section: Testing For Sars-cov-2 Antibodiesmentioning

confidence: 99%

Humoral Responses and Serological Assays in SARS-CoV-2 Infections

et al. 2020

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In December 2019, the novel betacoronavirus Severe Acute Respiratory Disease Coronavirus 2 (SARS-CoV-2) was first detected in Wuhan, China. SARS-CoV-2 has since become a pandemic virus resulting in hundreds of thousands of deaths and deep socioeconomic implications worldwide. In recent months, efforts have been directed towards detecting, tracking, and better understanding human humoral responses to SARS-CoV-2 infection. It has become critical to develop robust and reliable serological assays to characterize the abundance, neutralization efficiency, and duration of antibodies in virus-exposed individuals. Here we review the latest knowledge on humoral immune responses to SARS-CoV-2 infection, along with the benefits and limitations of currently available commercial and laboratory-based serological assays. We also highlight important serological considerations, such as antibody expression levels, stability and neutralization dynamics, as well as cross-reactivity and possible immunological back-boosting by seasonal coronaviruses. The ability to accurately detect, measure and characterize the various antibodies specific to SARS-CoV-2 is necessary for vaccine development, manage risk and exposure for healthcare and at-risk workers, and for monitoring reinfections with genetic variants and new strains of the virus. Having a thorough understanding of the benefits and cautions of standardized serological testing at a community level remains critically important in the design and implementation of future vaccination campaigns, epidemiological models of immunity, and public health measures that rely heavily on up-to-date knowledge of transmission dynamics.

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“…The characteristics of the study population and associated clinical specimens have previously been described. 8,12 In brief, the convalescent SARS-CoV-2 samples had been collected from COVID-19 patients who were confirmed positive by reverse transcription polymerase chain reaction (RT-PCR) at least 28 days prior to specimen collection (mean 45 days, SD 7.5). The pre-pandemic control samples were collected from a prior study of patients who presented to the Johns Hopkins Hospital Emergency Department with symptoms of an acute respiratory tract infection between January 2016 and June of 2019 as part of the Johns Hopkins Center for Influenza Research and Surveillance study.…”

Section: Clinical Specimens Testsmentioning

confidence: 99%

“…CoronaChek SARS-CoV-2 lateral flow assay and the Euroimmun IgG Spike ELISA were performed in accordance with the manufacturers' instructions and tests results previously reported on a different study on the same specimens. 8,12,13 Neutralizing antibody assay (titer and AUC determination) data were made available; their acquisition has previously been described. 12,14 Statistical Analyses Sensitivity of the assays was calculated among COVID-19 convalescent individuals and specificity was calculated among pre-pandemic population.…”

Section: Serological Assay Testing Using Commercial Assaysmentioning

confidence: 99%

“…To date, the rapid SARS-CoV-2 antibody tests approved under emergency use authorization are lateral flow assays, whose performance characteristics have been highly variable. 8 Furthermore, lateral flow tests do not offer any quantitative data on antibody levels; the latter are important given the wide range of antibody responses, particularly following mild SARS-CoV-2 infection. 9 We sought to develop a point-of-care test for SARS-CoV-2 antibody detection based on hemagglutination, leveraging a test platform that is already routinely used at the pointof-care for determination of blood types.…”

Section: Introductionmentioning

confidence: 99%

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A hemagglutination-based, semi-quantitative test for point-of-care determination of SARS-CoV-2 antibody levels

Kruse

Huang

Lee

et al. 2021

Preprint

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Serologic, point-of-care tests to detect antibodies against SARS-CoV-2 are an important tool in the COVID-19 pandemic. The majority of current point-of-care antibody tests developed for SARS-CoV-2 rely on lateral flow assays, but these do not offer quantitative information. To address this, we developed a new method of COVID-19 antibody testing employing hemagglutination tested on a dry card, similar to that which is already available for rapid typing of ABO blood groups. A fusion protein linking red blood cells (RBCs) to the receptor-binding domain (RBD) of SARS-CoV-2 spike protein was placed on the card. 200 COVID-19 patient and 200 control plasma samples were reconstituted with O-negative RBCs to form whole blood and added to the dried protein, followed by a stirring step and a tilting step, 3-minute incubation, and a second tilting step. The sensitivity for the hemagglutination test, Euroimmun IgG ELISA test and RBD-based CoronaChek lateral flow assay was 87.0%, 86.5%, and 84.5%, respectively, using samples obtained from recovered COVID-19 individuals. Testing pre-pandemic samples, the hemagglutination test had a specificity of 95.5%, compared to 97.3% and 98.9% for the ELISA and CoronaChek, respectively. A distribution of agglutination strengths was observed in COVID-19 convalescent plasma samples, with the highest agglutination score (4) exhibiting significantly higher neutralizing antibody titers than weak positives (2) (p<0.0001). Strong agglutinations were observed within 1 minute of testing, and this shorter assay time also increased specificity to 98.5%. In conclusion, we developed a novel rapid, point-of-care RBC agglutination test for the detection of SARS-CoV-2 antibodies that can yield semi-quantitative information on neutralizing antibody titer in patients. The five-minute test may find use in determination of serostatus prior to vaccination, post-vaccination surveillance and travel screening.

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Evaluation of Serological SARS-CoV-2 Lateral Flow Assays for Rapid Point of Care Testing

Cited by 21 publications

References 25 publications

Healthy donor T cell responses to common cold coronaviruses and SARS-CoV-2

Healthy donor T cell responses to common cold coronaviruses and SARS-CoV-2

Humoral Responses and Serological Assays in SARS-CoV-2 Infections

A hemagglutination-based, semi-quantitative test for point-of-care determination of SARS-CoV-2 antibody levels

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