Standardization of ELISA protocols for serosurveys of the SARS-CoV-2 pandemic using clinical and at-home blood sampling

Klumpp‐Thomas, Carleen; Kalish, Heather; Drew, Matthew; Hunsberger, Sally; Snead, Kelly; Fay, Michael P.; Mehalko, Jennifer; Shunmugavel, Anandakumar; Wall, Vanessa; Frank, Peter; Denson, John-Paul; Hong, Min Jee; Gulten, Gulcin; Messing, Simon; Hicks, Jennifer; Michael, Sam; Gillette, William; Hall, Matthew D.; Memoli, Matthew J.; Esposito, Dominic; Hall, Matthew D.

doi:10.1038/s41467-020-20383-x

Cited by 121 publications

(129 citation statements)

References 33 publications

(20 reference statements)

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“…Additionally, Spearman correlation matrices indicate significant associations (p < 0.005) between serum and DBS final interpreted results for IgG, IgM, IgA and nAb analytes (r = 0.872, r = 0.500, r = 0.781 and r = 0.814, respectively.). These findings are in line with similar studies [ 23–25 ]. Although DBS sampling does reduce sensitivity [ 8 , 26 ], our assay consistently demonstrated a similar level of sensitivity when the same samples were analyzed in serum.…”

Section: Resultssupporting

confidence: 94%

Validation of High-Throughput, Semiquantitative Solid-Phase Sarscoronavirus-2 Serology Assays in Serum and Dried Blood Spot Matrices

et al. 2021

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Aim: Serological assays for the detection of anti-SARS coronavirus-2 (SARS-CoV-2) antibodies are essential to the response to the global pandemic. A ligand binding-based serological assay was validated for the semiquantitative detection of IgG, IgM, IgA and neutralizing antibodies (nAb) against SARS-CoV-2 in serum. Results: The assay demonstrated high levels of diagnostic specificity and sensitivity (85–99% for all analytes). Serum IgG, IgM, IgA and nAb correlated positively (R2 = 0.937, R2 = 0.839, R2 = 0.939 and R2 = 0.501, p < 0.001, respectively) with those measured in dried blood spot samples collected using the hemaPEN® microsampling device (Trajan Scientific and Medical, Victoria, Australia). In vitro SARS-CoV-2 pseudotype neutralization correlated positively with the solid phase nAb signals in convalescent donors (R2 = 0.458, p < 0.05). Conclusion: The assay is applicable in efficacy studies, infection monitoring and postmarketing surveillance following vaccine rollout.

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Section: Resultssupporting

confidence: 94%

Validation of High-Throughput, Semiquantitative Solid-Phase Sarscoronavirus-2 Serology Assays in Serum and Dried Blood Spot Matrices

et al. 2021

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“…Eight 2-fold dilutions of sera prediluted in a ratio of 1:50 assay diluent were added to an equal volume of assay diluent (control) or to assay diluent mixes containing 5 μg/mL SARS-CoV-2 spike and 5 μg/mL RBD proteins (SARS-CoV-2 RBD-spike cocktail), or 5 μg/mL spike proteins from all 4 circulating coronaviruses (HKU1, OC43, 229E, NL63; circulating coronaviruses [cCoVs] cocktail) ( 25 ), for an on-plate assay dilution of 1:100 through 1:12,800. The dilution series was incubated for 30 minutes at room temperature and then analyzed using the multiplex antibody assay protocol, as mentioned previously.…”

Section: Methodsmentioning

confidence: 99%

A majority of uninfected adults show preexisting antibody reactivity against SARS-CoV-2

Majdoubi¹,

Michalski²,

O’Connell³

et al. 2021

JCI Insight

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Pre-existing cross-reactivity to SARS-CoV-2 may occur in absence of prior viral exposure. However, this has been difficult to quantify at the population level due to a lack of reliably defined seroreactivity thresholds. Using an orthogonal antibody testing approach, we estimated that 0.6% of non-triaged adults from the greater Vancouver area, Canada between May 17 th and June 19 th 2020 showed clear evidence of a prior SARS-CoV-2 infection, after adjusting for false-positive and false-negative test results. Using a highly sensitive multiplex assay and positive/negative thresholds established in infants in whom maternal antibodies have waned, we determine that more than 90% of uninfected adults showed antibody reactivity against the spike, receptor-binding domain (RBD), N-terminal domains (NTD) or the nucleocapsid (N) protein from SARS-CoV-2. This sero-reactivity was evenly distributed across age and sex, correlated with circulating coronaviruses reactivity, and was partially outcompeted by soluble circulating coronaviruses' spike. Using a custom SARS-CoV-2 peptide mapping array, we found that this antibody reactivity broadly mapped to spike, and to conserved non-structural viral proteins. We conclude that most adults display pre-existing antibody cross-reactivity against SARS-CoV-2, which further supports investigation of how this may impact the clinical severity of COVID-19 or SARS-CoV-2 vaccine responses.

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“…As with most respiratory viral diseases, it is difficult to estimate the true prevalence of the disease during a pandemic and the extent of its spread is only known after extensive study (1)(2)(3). The majority of patients infected with SARS-CoV-2 develop robust antibody responses against the viral spike protein, nucleocapsid protein, and the envelope protein that can be detected by CORONAVIRUS Undiagnosed SARS-CoV-2 seropositivity during the first six months of the COVID-19 pandemic in the United States serological testing (4)(5)(6)(7)(8). Antibodies against spike protein persist for months and can neutralize SARS-CoV-2 (9).…”

Section: Introductionmentioning

confidence: 99%

“…Protein and antigen sequences are available in (8). Statistical code is available at: https://zenodo.org/record/4958017#.YMkzYpNKh26.…”

mentioning

confidence: 99%

Undiagnosed SARS-CoV-2 seropositivity during the first 6 months of the COVID-19 pandemic in the United States

Kalish¹,

Klumpp‐Thomas²,

Hunsberger³

et al. 2021

Sci. Transl. Med.

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Asymptomatic SARS-CoV-2 infection and delayed implementation of diagnostics have led to poorly defined viral prevalence rates in the United States and elsewhere. To address this, we analyzed seropositivity in 9,089 adults in the United States who had not been diagnosed previously with COVID-19. Individuals with characteristics that reflected the US population (n = 27,716) were selected by quota sampling from 462,949 volunteers. Enrolled participants (n = 11,382) provided medical, geographic, demographic, and socioeconomic information, and dried blood samples. Survey questions coincident with the Behavioral Risk Factor Surveillance System survey, a large probability-based national survey, were used to adjust for selection bias. The majority (88.7%) of blood samples were collected between May 10th and July 31st, 2020 and were processed using ELISA to measure seropositivity (IgG and IgM antibodies against SARS-CoV-2 spike protein and the spike protein receptor binding domain). The overall weighted undiagnosed seropositivity estimate was 4.6% (95% CI: 2.6-6.5%) with race, age, sex, ethnicity, and urban/rural subgroup estimates ranging from 1.1% to 14.2%; the highest seropositivity estimates were in African American participants, younger, female, and Hispanic participants, and residents of urban centers. These data indicate that there were 4.8 undiagnosed SARS-CoV-2 infections for every diagnosed case of COVID-19, and an estimated 16.8 million infections were undiagnosed by mid-July 2020 in the United States.

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Standardization of ELISA protocols for serosurveys of the SARS-CoV-2 pandemic using clinical and at-home blood sampling

Cited by 121 publications

References 33 publications

Validation of High-Throughput, Semiquantitative Solid-Phase Sarscoronavirus-2 Serology Assays in Serum and Dried Blood Spot Matrices

Validation of High-Throughput, Semiquantitative Solid-Phase Sarscoronavirus-2 Serology Assays in Serum and Dried Blood Spot Matrices

A majority of uninfected adults show preexisting antibody reactivity against SARS-CoV-2

Undiagnosed SARS-CoV-2 seropositivity during the first 6 months of the COVID-19 pandemic in the United States

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