scholarly journals Diagnosis of Severe Acute Respiratory Syndrome (SARS) by Detection of SARS Coronavirus Nucleocapsid Antibodies in an Antigen-Capturing Enzyme-Linked Immunosorbent Assay

2003 ◽  
Vol 41 (12) ◽  
pp. 5781-5782 ◽  
Author(s):  
Y. Shi ◽  
Y. Yi ◽  
P. Li ◽  
T. Kuang ◽  
L. Li ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sars Coronavirus

scholarly journals Use of the COOH Portion of the Nucleocapsid Protein in an Antigen-Capturing Enzyme-Linked Immunosorbent Assay for Specific and Sensitive Detection of Severe Acute Respiratory Syndrome Coronavirus

2005 ◽  
Vol 12 (3) ◽  
pp. 474-476 ◽  
Author(s):  
Maofeng Qiu ◽  
Jin Wang ◽  
Hongxia Wang ◽  
Zeliang Chen ◽  
Erhei Dai ◽  
...  
Keyword(s):  
Amino Acids ◽  
Severe Acute Respiratory Syndrome ◽  
Epidemiological Study ◽  
Immunosorbent Assay ◽  
Nucleocapsid Protein ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sars Coronavirus ◽  
Specific Test ◽  
N Protein

ABSTRACT Antibody detection with a recombinant COOH portion of the severe acute respiratory syndrome (SARS) coronavirus nucleocapsid (N) protein, N13 (amino acids 221 to 422), was demonstrated to be more specific and sensitive than that with the full-length N protein, and an N13-based antigen-capturing enzyme-linked immunosorbent assay providing a convenient and specific test for serodiagnosis and epidemiological study of SARS was developed.

scholarly journals A Dual-Antigen Enzyme-Linked Immunosorbent Assay Allows the Assessment of Severe Acute Respiratory Syndrome Coronavirus 2 Antibody Seroprevalence in a Low-Transmission Setting

2020 ◽  
Vol 223 (1) ◽  
pp. 10-14 ◽  
Author(s):  
Sarah M Hicks ◽  
Kai Pohl ◽  
Teresa Neeman ◽  
Hayley A McNamara ◽  
Kate M Parsons ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Confidence Interval ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Elective Surgery ◽  
Enzyme Linked Immunosorbent Assay ◽  
Receptor Binding Domain ◽  
Assay Sensitivity ◽  
Specific Antibodies ◽  
Transmission Setting

Abstract Estimates of seroprevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies have been hampered by inadequate assay sensitivity and specificity. Using an enzyme-linked immunosorbent assay–based approach that combines data about immunoglobulin G responses to both the nucleocapsid and spike receptor binding domain antigens, we show that excellent sensitivity and specificity can be achieved. We used this assay to assess the frequency of virus-specific antibodies in a cohort of elective surgery patients in Australia and estimated seroprevalence in Australia to be 0.28% (95% Confidence Interval, 0–1.15%). These data confirm the low level of transmission of SARS-CoV-2 in Australia before July 2020 and validate the specificity of our assay.

scholarly journals Longitudinal Profile of Immunoglobulin G (IgG), IgM, and IgA Antibodies against the Severe Acute Respiratory Syndrome (SARS) Coronavirus Nucleocapsid Protein in Patients with Pneumonia Due to the SARS Coronavirus

2004 ◽  
Vol 11 (4) ◽  
pp. 665-668 ◽  
Author(s):  
Patrick C. Y. Woo ◽  
Susanna K. P. Lau ◽  
Beatrice H. L. Wong ◽  
Kwok-hung Chan ◽  
Chung-ming Chu ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Indirect Immunofluorescence ◽  
Immunoglobulin G ◽  
Nucleocapsid Protein ◽  
Baseline Level ◽  
Immunofluorescence Assay ◽  
Indirect Immunofluorescence Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sars Coronavirus ◽  
Iga Antibodies

ABSTRACT By using a recombinant severe acute respiratory syndrome coronavirus (SARS-CoV) nucleocapsid protein-based enzyme-linked immunosorbent assay (ELISA) and serum specimens serially collected (from day 0 to day 240 after symptom onset) from patients with pneumonia due to SARS-CoV, we analyzed the longitudinal profiles of immunoglobulin G (IgG), IgM, and IgA antibodies against the SARS-CoV nucleocapsid protein in patients with pneumonia due to SARS-CoV. For IgG, the median optical density at 450 nm (OD450) turned positive at day 17 and a biphasic response was observed. At day 240, all patients were still positive for anti-nucleocapsid protein IgG antibody. For IgM, the median OD450 turned positive at day 20.5, peaked at about day 80, and fell to below the baseline level at about day 180. At day 240, 36% of the patients were still positive for anti-nucleocapsid protein IgM antibody. For IgA, the median OD450 turned positive at day 17, peaked at about day 50, and fell to below the baseline level at about day 180. At day 240, 36% of the patients were still positive for anti-nucleocapsid protein IgA antibody. The time of seroconversion detected by the recombinant SARS-CoV nucleocapsid protein-based ELISA and that detected by indirect immunofluorescence assay were similar. The median times of seroconversion for IgG, IgM, and IgA detected by the indirect immunofluorescence assay were 17 days (17 days by ELISA), 16.5 days (20.5 days by ELISA), and 17.5 days (17 days by ELISA), respectively, after disease onset. One, four, and one of the six patients who died did not produce any IgG, IgM, and IgA antibodies against the nucleocapsid protein of SARS-CoV, respectively, although these antibodies were detected in all six patients by the indirect immunofluorescence assay. Further studies should be performed to see whether SARS-CoV nucleocapsid protein antibody positivity has any prognostic significance.

scholarly journals The landscape of antibody binding in SARS-CoV-2 infection

2020 ◽  
Author(s):  
Anna S Heffron ◽  
Sean J McIlwain ◽  
Maya F Amjadi ◽  
David A Baker ◽  
Saniya Khullar ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Diagnostic Accuracy ◽  
B Cell ◽  
Immunosorbent Assay ◽  
Antibody Binding ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Peptide Microarray ◽  
B Cell Epitopes ◽  
Homologous Peptide

The search for potential antibody-based diagnostics, vaccines, and therapeutics for pandemic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has focused almost exclusively on the spike (S) and nucleocapsid (N) proteins. Coronavirus membrane (M), ORF3a, and ORF8 proteins are humoral immunogens in other coronaviruses (CoVs) but remain largely uninvestigated for SARS-CoV-2. Here we use ultradense peptide microarray mapping to show that SARS-CoV-2 infection induces robust antibody responses to epitopes throughout the SARS-CoV-2 proteome, particularly in M, in which one epitope achieved excellent diagnostic accuracy. We map 79 B cell epitopes throughout the SARS-CoV-2 proteome and demonstrate that antibodies that develop in response to SARS-CoV-2 infection bind homologous peptide sequences in the six other known human CoVs. We also confirm reactivity against four of our top-ranking epitopes by enzyme-linked immunosorbent assay (ELISA). Illness severity correlated with increased reactivity to nine SARS-CoV-2 epitopes in S, M, N, and ORF3a in our population. Our results demonstrate previously unknown, highly reactive B cell epitopes throughout the full proteome of SARS-CoV-2 and other CoV proteins.

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

Vaccines ◽  
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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