From multiplex serology to serolomics – a novel approach to the antibody response against the SARS-CoV-2 proteome

Butt, Julia; Murugan, Rajagopal; Hippchen, Theresa; Olberg, Sylvia; Straaten, Monique van; Wardemann, Hedda; Stebbins, Erec; Kräusslich, Hans‐Georg; Bartenschlager, Ralf; Brugger, Hermann; Laketa, Vibor; Schöttker, Ben; Müller, Bárbara; Merle, Uta; Waterboer, Tim

doi:10.1101/2020.10.19.20214916

Cited by 3 publications

(6 citation statements)

References 31 publications

(52 reference statements)

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“…Studies by our team and others found a slightly lower specificity when including borderline results (eg, 96.3% or 98.6%). 16,26 Given the low SARS-CoV-2 prevalence in the general population, when this study was performed, test combinations with specificities greater than 99% are required to obtain valid results. A combination of 2 different tests improves accuracy markedly and has recently been used in a large seroepidemiological study.…”

Section: Discussionmentioning

confidence: 99%

“…15 Here, we combined ELISA with immunofluorescence as complementary tests for all samples. 16 Moreover, we further analyzed discordant samples by additional tests to maximize both specificity and sensitivity. As shown in Table 2, both initial assays independently yielded 40% to 50% of false-positive results, but these could be eliminated by a combination of the 2 readouts.…”

Section: Discussionmentioning

confidence: 99%

“…Discordant results were clarified by electrochemiluminescence immunoassays, a second ELISA, or an in-house Luminex-based assay. We had previously determined that a combination of these complementary assays considerably increases the specificity of the results without compromising the sensitivity. The workflow of the serological analyses is shown eTable 1 in the Supplement ; details of the serological analyses, the immunofluorescence assay, and the neutralization assay are given in the eMethods in the Supplement .…”

Section: Methodsmentioning

confidence: 99%

“…A number of samples (n = 119) displayed borderline reactivity in ELISA testing or yielded discordant results between ELISA and immunofluorescence tests. 16 In these cases, at least 1 of the additional tests was used for final classification. As shown in…”

Section: Detection Of Acute Sars-cov-2 Infection By Rt-pcrmentioning

confidence: 99%

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Prevalence of SARS-CoV-2 Infection in Children and Their Parents in Southwest Germany

et al. 2021

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daycare closures were enforced as measures to confine the novel coronavirus disease 2019 (COVID-19) pandemic, based on the assumption that young children may play a key role in severe acute respiratory coronavirus 2 (SARS-CoV-2) spread. Given the grave consequences of contact restrictions for children, a better understanding of their contribution to the COVID-19 pandemic is of great importance. OBJECTIVE To describe the rate of SARS-CoV-2 infections and the seroprevalence of SARS-CoV-2 antibodies in children aged 1 to 10 years, compared with a corresponding parent of each child, in a population-based sample. DESIGN, SETTING, AND PARTICIPANTS This large-scale, multicenter, cross-sectional investigation (the COVID-19 BaWü study) enrolled children aged 1 to 10 years and a corresponding parent between April 22 and May 15, 2020, in southwest Germany. EXPOSURES Potential exposure to SARS-CoV-2. MAIN OUTCOMES AND MEASURES The main outcomes were infection and seroprevalence of SARS-CoV-2. Participants were tested for SARS-CoV-2 RNA from nasopharyngeal swabs by reverse transcription-polymerase chain reaction and SARS-CoV-2 specific IgG antibodies in serum by enzyme-linked immunosorbent assays and immunofluorescence tests. Discordant results were clarified by electrochemiluminescence immunoassays, a second enzyme-linked immunosorbent assay, or an in-house Luminex-based assay. RESULTS This study included 4964 participants: 2482 children (median age, 6 [range, 1-10] years; 1265 boys [51.0%]) and 2482 parents (median age, 40 [range, 23-66] years; 615 men [24.8%]). Two participants (0.04%) tested positive for SARS-CoV-2 RNA. The estimated SARS-CoV-2 seroprevalence was low in parents (1.8% [95% CI, 1.2-2.4%]) and 3-fold lower in children (0.6% [95% CI, 0.3-1.0%]). Among 56 families with at least 1 child or parent with seropositivity, the combination of a parent with seropositivity and a corresponding child with seronegativity was 4.3 (95% CI, 1.19-15.52) times higher than the combination of a parent who was seronegative and a corresponding child with seropositivity. We observed virus-neutralizing activity for 66 of 70 IgG-positive serum samples (94.3%). CONCLUSIONS AND RELEVANCE In this cross-sectional study, the spread of SARS-CoV-2 infection during a period of lockdown in southwest Germany was particularly low in children aged 1 to 10 years. Accordingly, it is unlikely that children have boosted the pandemic. This SARS-CoV-2 prevalence study, which appears to be the largest focusing on children, is instructive for how ad hoc mass testing provides the basis for rational political decision-making in a pandemic.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Detection Of Acute Sars-cov-2 Infection By Rt-pcrmentioning

confidence: 99%

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Prevalence of SARS-CoV-2 Infection in Children and Their Parents in Southwest Germany

et al. 2021

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“…Immunoglobulin (Ig) G (IgG) antibody levels against the S1 protein of SARS-CoV-2 were determined using an enzyme-linked immunosorbent assay (ELISA) kit (EI 2606–9601 G; Euroimmun AG) run on an Analyzer I instrument (Euroimmun AG). Multiplex SARS-CoV-2 serology was performed as recently described, and briefly, N and S1-RBD antibody levels measured in median fluorescence intensity (MFI) units [ 11 ]. Competition of sera with S1-ACE2 interaction was measured using the cPass surrogate SARS-CoV-2 neutralization test kit (L00847; Genscript, Piscataway, NJ, USA).…”

Section: Methodsmentioning

confidence: 99%

Persistent Symptoms in Adult Patients 1 Year After Coronavirus Disease 2019 (COVID-19): A Prospective Cohort Study

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Waterboer

Hippchen

et al. 2021

Clinical Infectious Diseases

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Background Long COVID is defined as the persistence of symptoms beyond 3 months after SARS-CoV-2 infection. To better understand the long-term course and etiology of symptoms we analyzed a cohort of COVID-19 patients prospectively. Methods Patients were included at 5 months after acute COVID-19 in this prospective, non-interventional follow-up study. Patients followed until 12 months after COVID-19 symptom onset (n=96, 32.3% hospitalised, 55.2% females) were included in this analysis of symptoms, quality of life (based on a SF-12 survey), laboratory parameters including antinuclear antibodies (ANA), and SARS-CoV-2 antibody levels. Results At month 12, only 22.9% of patients were completely free of symptoms and the most frequent symptoms were reduced exercise capacity (56.3%), fatigue (53.1%), dyspnoea (37.5%), concentration problems (39.6%), problems finding words (32.3%), and sleeping problems (26.0%). Females showed significantly more neurocognitive symptoms than males. ANA titres were ≥1:160 in 43.6% of patients at 12 months post COVID-19 symptom onset, and neurocognitive symptom frequency was significantly higher in the group with an ANA titre ≥1:160 compared to <1:160. Compared to patients without symptoms, patients with at least one long COVID symptom at 12 months did not differ significantly with respect to their SARS-CoV-2-antibody levels, but had a significantly reduced physical and mental life quality compared to patients without symptoms. Conclusions Neurocognitive long COVID symptoms can persist at least for one year after COVID-19 symptom onset, and reduce life quality significantly. Several neurocognitive symptoms were associated with ANA titre elevations. This may indicate autoimmunity as cofactor in aetiology of long COVID.

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Single‐reaction multi‐antigen serological test for comprehensive evaluation of SARS‐CoV‐2 patients by flow cytometry

et al. 2021

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Here, we describe a new, simple, highly multiplexed serological test that generates a more complete picture of seroconversion than single antigen‐based assays. Flow cytometry is used to detect multiple Ig isotypes binding to four SARS‐CoV‐2 antigens: the Spike glycoprotein, its RBD fragment (the main target for neutralizing antibodies), the nucleocapsid protein, and the main cysteine‐like protease in a single reaction. Until now, most diagnostic serological tests measured antibodies to only one antigen and in some laboratory‐confirmed patients no SARS‐CoV‐2‐specific antibodies could be detected. Our data reveal that while most patients respond against all the viral antigens tested, others show a marked bias to make antibodies against either proteins exposed on the viral particle or those released after cellular infection. With this assay, it was possible to discriminate between patients and healthy controls with 100% confidence. Analysing the response of multiple Ig isotypes to the four antigens in combination may also help to establish a correlation with the severity degree of disease. A more detailed description of the immune responses of different patients to SARS‐CoV‐2 virus might provide insight into the wide array of clinical presentations of COVID‐19.

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From multiplex serology to serolomics – a novel approach to the antibody response against the SARS-CoV-2 proteome

Cited by 3 publications

References 31 publications

Prevalence of SARS-CoV-2 Infection in Children and Their Parents in Southwest Germany

Prevalence of SARS-CoV-2 Infection in Children and Their Parents in Southwest Germany

Persistent Symptoms in Adult Patients 1 Year After Coronavirus Disease 2019 (COVID-19): A Prospective Cohort Study

Single‐reaction multi‐antigen serological test for comprehensive evaluation of SARS‐CoV‐2 patients by flow cytometry

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