Evaluation of the EUROIMMUN Anti-SARS-CoV-2 ELISA Assay for detection of IgA and IgG antibodies

AbstractAs the Coronavirus 2019 (COVID-19) pandemic evolves, the development of immunoassays to help determine exposure and potentially predict immunity has become a pressing priority. In this report we present the performance of the EUROIMMUN enzyme-linked immunosorbent assay (ELISA) for semi-quantitative detection of IgA and IgG antibodies in serum and plasma samples using recombinant S1 domain of the SARS-CoV-2 spike protein as antigen. Specimens from patients, with and without COVID-19 infection, were tested at the University of Chicago Clinical Microbiology and Immunology Laboratory. Of 57 samples from COVID-19 PCR-negative patients, including 28 samples positive for common human coronavirus strains, 53 tested negative and 4 tested positive for IgA (93.0% agreement) while 56 tested negative and 1 tested positive for IgG (98.2% agreement). For COVID-19 PCR-positive patients, 29 of 30 (96.7%) samples collected ≥3 days after positive PCR were positive for IgA, and 28 of 28 samples collected ≥4 days after positive PCR were positive for IgG.The EUROIMMUN Anti-SARS-CoV-2 ELISA Assay demonstrates excellent sensitivity for detection of IgA and IgG antibodies from samples collected ≥3 days and ≥4 days, respectively, after COVID-19 diagnosis by PCR. This assay did not demonstrate cross reaction in any of the 28 samples from patients with common human coronaviruses, including types HKU1, NL63, CV229E, and OC43.

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Performance of the EUROIMMUN Anti-SARS-CoV-2 ELISA Assay for detection of IgA and IgG antibodies in South Africa

PLoS ONE ◽

10.1371/journal.pone.0252317 ◽

2021 ◽

Vol 16 (6) ◽

pp. e0252317

Author(s):

Maemu P. Gededzha ◽

Nakampe Mampeule ◽

Sarika Jugwanth ◽

Nontobeko Zwane ◽

Anura David ◽

...

Keyword(s):

South Africa ◽

Diagnostic Algorithm ◽

Clinical Syndrome ◽

Quantitative Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Elisa Assay ◽

Igg Antibodies ◽

Assay Sensitivity ◽

Pcr Diagnosis ◽

Negative Controls

Severe Acute Respiratory Syndrome-Coronavirus 2 (SARS-CoV-2) has been identified as the causative agent for causing the clinical syndrome of COVID -19. Accurate detection of SARS-CoV-2 infection is not only important for management of infected individuals but also to break the chain of transmission. South Africa is the current epicenter of SARS-CoV-2 infection in Africa. To optimize the diagnostic algorithm for SARS-CoV-2 in the South African setting, the study aims to evaluate the diagnostic performance of the EUROIMMUN Anti-SARS-CoV-2 assays. This study reported the performance of EUROIMMUN enzyme-linked immunosorbent assay (ELISA) for semi-quantitative detection of IgA and IgG antibodies in serum and plasma samples targeting the recombinant S1 domain of the SARS-CoV-2 spike protein as antigen. Samples were collected from 391 individuals who had tested positive for SARS-CoV-2 and 139 SARS CoV-2 negative controls. Samples were stratified by number of days’ post-PCR diagnosis and symptoms. The sensitivity of EUROIMMUN IgG was 64.1% (95% CI: 59.1–69.0%) and 74.3% (95% CI: 69.6–78.6%) for IgA and the specificity was lower for IgA [84.2% (95% CI: 77–89.2%)] than IgG [95.2% (95% CI: 90.8–98.4%)]. The EUROIMMUN Anti-SARS-CoV-2 ELISA Assay sensitivity was higher for IgA but low for IgG and improved for both assays in symptomatic individuals and at later timepoints post PCR diagnosis.

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Specific serology for emerging human coronaviruses by protein microarray

Eurosurveillance ◽

10.2807/1560-7917.es2013.18.14.20441 ◽

2013 ◽

Vol 18 (14) ◽

Cited By ~ 62

Author(s):

C Reusken ◽

H Mou ◽

G J Godeke ◽

L van der Hoek ◽

B Meyer ◽

...

Keyword(s):

Receptor Binding ◽

Protein Microarray ◽

Spike Protein ◽

Microarray Technology ◽

Igg Antibodies ◽

Specific Detection ◽

Human Coronavirus ◽

Serological Assay ◽

Human Coronaviruses ◽

Binding Subunit

We present a serological assay for the specific detection of IgM and IgG antibodies against the emerging human coronavirus hCoV-EMC and the SARS-CoV based on protein microarray technology. The assay uses the S1 receptor-binding subunit of the spike protein of hCoV-EMC and SARS-CoV as antigens. The assay has been validated extensively using putative cross-reacting sera of patient cohorts exposed to the four common hCoVs and sera from convalescent patients infected with hCoV-EMC or SARS-CoV.

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A Quantitative ELISA Protocol for Detection of Specific Human IgG Against the SARS-CoV-2 Spike Protein

Vaccines ◽

10.3390/vaccines9070770 ◽

2021 ◽

Vol 9 (7) ◽

pp. 770

Author(s):

Rémi Vernet ◽

Emily Charrier ◽

Julien Grogg ◽

Nicolas Mach

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Immunosorbent Assay ◽

Spike Protein ◽

Quantitative Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibodies ◽

Human Igg ◽

Microtiter Plates ◽

Global Priority ◽

Humoral Responses

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a worldwide pandemic with at least 3.8 million deaths to date. For that reason, finding an efficient vaccine for this virus quickly became a global priority. The majority of vaccines now marketed are based on the SARS‑CoV‑2 spike protein that has been described as the keystone for optimal immunization. In order to monitor SARS‑CoV‑2 spike-specific humoral responses generated by immunization or infection, we have developed a robust and reproducible enzyme-linked immunosorbent assay (ELISA) protocol. This protocol describes a method for quantitative detection of IgG antibodies against the SARS‑CoV‑2 spike protein using antigen-coated microtiter plates. Results showed that antibodies could be quantified between the range of 1.953 ng/mL to 500 ng/mL with limited inter- and intra-assay variability.

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Evaluation of Enzyme-Linked Immunoassay and Colloidal Gold-Immunochromatographic Assay Kit for Detection of Novel Coronavirus (SARS-Cov-2) Causing an Outbreak of Pneumonia (COVID-19)

10.1101/2020.02.27.20028787 ◽

2020 ◽

Cited By ~ 42

Author(s):

Jie Xiang ◽

Mingzhe Yan ◽

Hongze Li ◽

Ting Liu ◽

Chenyao Lin ◽

...

Keyword(s):

Colloidal Gold ◽

Laboratory Diagnosis ◽

Immunochromatographic Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Healthy Controls ◽

Plasma Samples ◽

Igg Antibodies ◽

Significant Difference ◽

Novel Coronavirus ◽

Treatment Pressure

AbstractBACKGROUNDIn December 2019, a novel coronavirus (SARS-CoV-2)–infected pneumonia (COVID-19) occurred in Wuhan, China. Travel-associated cases have also been reported in other countries. The number of cases has increased rapidly but laboratory diagnosis is limited.METHODSWe collect two groups of cases diagnosed with COVID-19 for experiments. One group collected 63 samples for Enzyme-linked immunosorbent assay (ELISA) IgG and IgM antibodies. The other group collected 91 plasma samples for colloidal gold-immunochromatographic assay (GICA).RESULTSThe sensitivity of the combined ELISA IgM and ELISA IgG detection was 55/63 ( 87.3%), The sensitivity of the combined GICA IgM and GICA IgG detection was 75/91 ( 82.4%), Both methods are negative for healthy controls, specificity of 100%. There is no significant difference between the sensitivity of between ELISA and GICA (IgM+ IgG).CONCLUSIONSELISA and GICA for specific IgM and IgG antibodies are conventional serological assays, they are simple, fast, and safe, the results can be used for clinical reference, and the huge clinical diagnosis and treatment pressure can be greatly relieved.

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Development of a Bead-Based Multiplex Immunoassay for Simultaneous Quantitative Detection of IgG Serum Antibodies against Measles, Mumps, Rubella, and Varicella-Zoster Virus

Clinical and Vaccine Immunology ◽

10.1128/cvi.05537-11 ◽

2012 ◽

Vol 19 (3) ◽

pp. 396-400 ◽

Cited By ~ 59

Author(s):

Gaby P. Smits ◽

Pieter G. van Gageldonk ◽

Leo M. Schouls ◽

Fiona R. M. van der Klis ◽

Guy A. M. Berbers

Keyword(s):

Varicella Zoster Virus ◽

High Specificity ◽

Good Alternative ◽

Multiplex Assay ◽

Quantitative Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibodies ◽

Multiplex Immunoassay ◽

Varicella Zoster ◽

Elisa Kit

ABSTRACTEnzyme-linked immunosorbent assay (ELISA) is normally used to quantify the amount of serum IgG antibodies against measles, mumps, rubella, and varicella-zoster virus (MMRV). However, this method is time- and material-consuming. Therefore, a multiplex immunoassay for the simultaneous quantitative detection of antibodies against MMRV was developed. In-house as well as commercially available antigens can be used, making the assay available for all laboratories. The multiplex assay is much more sensitive than the separate ELISAs and has a high specificity, and only 5 μl of serum is needed. Heterologous inhibition did not exceed 11.5%, while homologous inhibition varied between 91.3 and 97.9%. Good correlations with the in-house ELISAs for measles (R2= 0.98), mumps (R2= 0.97), and rubella (R2= 0.97) virus as well as with the ELISA kit for varicella-zoster virus (R2= 0.95) were obtained. In conclusion, the MMRV multiplex assay is a good alternative to the conventional ELISAs and suitable for use in serosurveillance and vaccine studies.

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Evaluation of the IgG antibody response to SARS CoV-2 infection and performance of a lateral flow immunoassay: cross-sectional and longitudinal analysis over 11 months

BMJ Open ◽

10.1136/bmjopen-2020-048142 ◽

2021 ◽

Vol 11 (6) ◽

pp. e048142

Author(s):

Louise J Robertson ◽

Julie S Moore ◽

Kevin Blighe ◽

Kok Yew Ng ◽

Nigel Quinn ◽

...

Keyword(s):

Northern Ireland ◽

Post Infection ◽

Rapid Test ◽

Spike Protein ◽

Lateral Flow Immunoassay ◽

Plasma Samples ◽

Igg Antibodies ◽

Igg Antibody ◽

The Uk ◽

Antibody Levels

ObjectiveTo evaluate the dynamics and longevity of the humoral immune response to SARS-CoV-2 infection and assess the performance of professional use of the UK-RTC AbC-19 Rapid Test lateral flow immunoassay (LFIA) for the target condition of SARS-CoV-2 spike protein IgG antibodies.DesignNationwide serological study.SettingNorthern Ireland, UK, May 2020–February 2021.ParticipantsPlasma samples were collected from a diverse cohort of individuals from the general public (n=279), Northern Ireland healthcare workers (n=195), pre-pandemic blood donations and research studies (n=223) and through a convalescent plasma programme (n=183). Plasma donors (n=101) were followed with sequential samples over 11 months post-symptom onset.Main outcome measuresSARS-CoV-2 antibody levels in plasma samples using Roche Elecsys Anti-SARS-CoV-2 IgG/IgA/IgM, Abbott SARS-CoV-2 IgG and EuroImmun IgG SARS-CoV-2 ELISA immunoassays over time. UK-RTC AbC-19 LFIA sensitivity and specificity, estimated using a three-reference standard system to establish a characterised panel of 330 positive and 488 negative SARS-CoV-2 IgG samples.ResultsWe detected persistence of SARS-CoV-2 IgG antibodies for up to 10 months post-infection, across a minimum of two laboratory immunoassays. On the known positive cohort, the UK-RTC AbC-19 LFIA showed a sensitivity of 97.58% (95.28% to 98.95%) and on known negatives, showed specificity of 99.59% (98.53 % to 99.95%).ConclusionsThrough comprehensive analysis of a cohort of pre-pandemic and pandemic individuals, we show detectable levels of IgG antibodies, lasting over 46 weeks when assessed by EuroImmun ELISA, providing insight to antibody levels at later time points post-infection. We show good laboratory validation performance metrics for the AbC-19 rapid test for SARS-CoV-2 spike protein IgG antibody detection in a laboratory-based setting.

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Differential Downregulation of ACE2 by the Spike Proteins of Severe Acute Respiratory Syndrome Coronavirus and Human Coronavirus NL63

Journal of Virology ◽

10.1128/jvi.01248-09 ◽

2009 ◽

Vol 84 (2) ◽

pp. 1198-1205 ◽

Cited By ~ 176

Author(s):

Ilona Glowacka ◽

Stephanie Bertram ◽

Petra Herzog ◽

Susanne Pfefferle ◽

Imke Steffen ◽

...

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Lung Injury ◽

Vero Cells ◽

Cell Entry ◽

Spike Protein ◽

Converting Enzyme ◽

Human Coronavirus ◽

Angiotensin Converting Enzyme 2 ◽

Similar Activity ◽

Human Coronaviruses

ABSTRACT The human coronaviruses (CoVs) severe acute respiratory syndrome (SARS)-CoV and NL63 employ angiotensin-converting enzyme 2 (ACE2) for cell entry. It was shown that recombinant SARS-CoV spike protein (SARS-S) downregulates ACE2 expression and thereby promotes lung injury. Whether NL63-S exerts a similar activity is yet unknown. We found that recombinant SARS-S bound to ACE2 and induced ACE2 shedding with higher efficiency than NL63-S. Shedding most likely accounted for the previously observed ACE2 downregulation but was dispensable for viral replication. Finally, SARS-CoV but not NL63 replicated efficiently in ACE2-positive Vero cells and reduced ACE2 expression, indicating robust receptor interference in the context of SARS-CoV but not NL63 infection.

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Evaluation of commercially available immuno-magnetic agglutination and enzyme-linked immunosorbent assays for rapid point-of-care diagnostics of COVID-19

10.1101/2020.08.15.20172080 ◽

2020 ◽

Author(s):

Maria Engel Moeller ◽

Jeppe Fock ◽

Pearlyn Pah ◽

Antia De La Campa Veras ◽

Melanie Bade ◽

...

Keyword(s):

Point Of Care ◽

Enzyme Linked Immunosorbent Assay ◽

Plasma Samples ◽

Igg Antibodies ◽

Chain Reaction ◽

Point Of Care Test ◽

Point Of Care Diagnostics ◽

Polymerase Chain ◽

Respiratory Coronavirus ◽

Immunomagnetic Assay

Introduction: Coronavirus Disease 2019 (COVID-19) is caused by severe acute respiratory coronavirus-2 (SARS-CoV-2). Fast, accurate and simple blood-based assays for quantification of anti-SARS-CoV-2 antibodies are urgently needed to identify infected individuals and keep track of the spread of disease. Methods: The study included 35 plasma samples from 22 individuals with confirmed COVID-19 by real time reverse transcriptase polymerase chain reaction and 40 non COVID-19 plasma samples. Anti-SARS-CoV-2 IgM/IgA or IgG antibodies were detected by a microfluidic quantitative immunomagnetic assay (IMA)(ViroTrack Sero COVID IgM+IgA/IgG Ab, Blusense Diagnostics, Denmark) and by enzyme-linked immunosorbent assay ((ELISA) (EuroImmun Medizinische Labordiagnostika, Germany). Results: Of the 35 plasma samples from the COVID-19 patients, 29 (82.9%) were positive for IgA/IgM or IgG by IMA and 29 samples (82.9%) were positive by ELISA. Sensitivity for only one sample per patient was 68% for IgA+IgM and 73% IgG by IMA and 73% by ELISA. For samples collected 14 days after symptom onset, the sensitivity of both IMA and ELISA was around 90%. Specificity of the IMA reached 100% compared to 95% for ELISA IgA and 97.5% for ELISA IgG. Conclusion: IMA for COVID-19 is a rapid simple-to-use point of care test with sensitivity and specificity similar to a commercial ELISA.

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Evaluation of Nucleocapsid and Spike Protein-Based Enzyme-Linked Immunosorbent Assays for Detecting Antibodies against SARS-CoV-2

Journal of Clinical Microbiology ◽

10.1128/jcm.00461-20 ◽

2020 ◽

Vol 58 (6) ◽

Cited By ~ 140

Author(s):

Wanbing Liu ◽

Lei Liu ◽

Guomei Kou ◽

Yaqiong Zheng ◽

Yinjuan Ding ◽

...

Keyword(s):

Nucleocapsid Protein ◽

Early Stage ◽

Disease Onset ◽

High Sensitivity ◽

Spike Protein ◽

Nucleic Acid Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibodies ◽

Serum Samples ◽

Positive Rate

ABSTRACT At present, PCR-based nucleic acid detection cannot meet the demands for coronavirus infectious disease (COVID-19) diagnosis. Two hundred fourteen confirmed COVID-19 patients who were hospitalized in the General Hospital of Central Theater Command of the People’s Liberation Army between 18 January and 26 February 2020 were recruited. Two enzyme-linked immunosorbent assay (ELISA) kits based on recombinant severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid protein (rN) and spike protein (rS) were used for detecting IgM and IgG antibodies, and their diagnostic feasibility was evaluated. Among the 214 patients, 146 (68.2%) and 150 (70.1%) were successfully diagnosed with the rN-based IgM and IgG ELISAs, respectively; 165 (77.1%) and 159 (74.3%) were successfully diagnosed with the rS-based IgM and IgG ELISAs, respectively. The positive rates of the rN-based and rS-based ELISAs for antibody (IgM and/or IgG) detection were 80.4% and 82.2%, respectively. The sensitivity of the rS-based ELISA for IgM detection was significantly higher than that of the rN-based ELISA. We observed an increase in the positive rate for IgM and IgG with an increasing number of days post-disease onset (d.p.o.), but the positive rate of IgM dropped after 35 d.p.o. The positive rate of rN-based and rS-based IgM and IgG ELISAs was less than 60% during the early stage of the illness, 0 to 10 d.p.o., and that of IgM and IgG was obviously increased after 10 d.p.o. ELISA has a high sensitivity, especially for the detection of serum samples from patients after 10 d.p.o., so it could be an important supplementary method for COVID-19 diagnosis.

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Evaluation of Nucleocapsid and Spike Protein-based ELISAs for detecting antibodies against SARS-CoV-2

10.1101/2020.03.16.20035014 ◽

2020 ◽

Cited By ~ 14

Author(s):

Wanbing Liu ◽

Lei Liu ◽

Guomei Kou ◽

Yaqiong Zheng ◽

Yinjuan Ding ◽

...

Keyword(s):

Nucleocapsid Protein ◽

Early Stage ◽

Disease Onset ◽

High Sensitivity ◽

Spike Protein ◽

Nucleic Acid Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibodies ◽

Serum Samples ◽

Positive Rate

AbstractBackgroundAt present, PCR-based nucleic acid detection cannot meet the demands for coronavirus infectious disease (COVID-19) diagnosis.Methods214 confirmed COVID-19 patients who were hospitalized in the General Hospital of Central Theater Command of the People’s Liberation Army between January 18 and February 26, 2020, were recruited. Two Enzyme-Linked Immunosorbent Assay (ELISA) kits based on recombinant SARS-CoV-2 nucleocapsid protein (rN) and spike protein (rS) were used for detecting IgM and IgG antibodies, and their diagnostic feasibility was evaluated.ResultsAmong the 214 patients, 146 (68.2%) and 150 (70.1%) were successfully diagnosed with the rN-based IgM and IgG ELISAs, respectively; 165 (77.1%) and 159 (74.3%) were successfully diagnosed with the rS-based IgM and IgG ELISAs, respectively. The positive rates of the rN-based and rS-based ELISAs for antibody (IgM and/or IgG) detection were 80.4% and 82.2%, respectively. The sensitivity of the rS-based ELISA for IgM detection was significantly higher than that of the rN-based ELISA. We observed an increase in the positive rate for IgM and IgG with an increasing number of days post-disease onset (d.p.o.), but the positive rate of IgM dropped after 35 d.p.o. The positive rate of rN-based and rS-based IgM and IgG ELISAs was less than 60% during the early stage of the illness 0-10 d.p.o., and that of IgM and IgG was obviously increased after 10 d.p.o.ConclusionsELISA has a high sensitivity, especially for the detection of serum samples from patients after 10 d.p.o, it can be an important supplementary method for COVID-19 diagnosis.

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