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

Liu, Wanbing; Liu, Lei; Kou, Guomei; Zheng, Yaqiong; Ding, Yinjuan; Ni, Wenxu; Wang, Qiongshu; Tan, Li; Wu, Wanlei; Tang, Shi; Xiong, Zhuo; Zheng, Shangen

doi:10.1128/jcm.00461-20

Cited by 597 publications

(571 citation statements)

References 17 publications

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“…Initial reports suggest that following infection with SARS-CoV-2, the immune system takes 6-21 days to product IgM and IgG antibodies [5,7,17,18]. Indeed, the RDT manufacturer states that antibodies only become detectable 7 days after disease onset.…”

Section: Discussionmentioning

confidence: 99%

Sensitivity and specificity of a rapid test for assessment of exposure to SARS-CoV-2 in a community-based setting in Brazil

Pellanda

2020

Preprint

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Background: While the recommended laboratory diagnosis of COVID-19 is a molecular based assay, population-based studies to determine the prevalence of COVID-19 usually use serological assays. Objective: To evaluate the sensitivity and specificity of a rapid diagnostic test for COVID-19 compared to quantitative reverse transcription polymerase chain reaction (qRT-PCR). Methods: We evaluated the sensitivity using a panel of finger prick blood samples from participants >18 years of age that had been tested for COVID-19 by qRT-PCR. For assessing specificity, we used serum samples from the 1982 Pelotas (Brazil) Birth Cohort participants collected in 2012 with no exposure to SARS-CoV-2. Results: The sensitivity of the test was 77.1% (95% CI 66.6 -85.6), based upon 83 subjects who had tested positive for qRT-PCR at least 10 days before the rapid diagnostic test (RDT). Based upon 100 sera samples, specificity was 98.0% (95% CI 92.9 -99.8). There was substantial agreement (Kappa score 0.76) between the qRT-PCR results and the RDT. Interpretation. The validation results are well in line with previous assessments of the test, and confirm that it is sufficiently precise for epidemiological studies aimed at monitoring levels and trends of the COVID-19 pandemic.

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Section: Discussionmentioning

confidence: 99%

Sensitivity and specificity of a rapid test for assessment of exposure to SARS-CoV-2 in a community-based setting in Brazil

Pellanda

2020

Preprint

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“…The number of lymphocytes was significantly reduced and C-reactive protein (CRP) was significantly increased in severe cases [10][11][12][13]. During the submission of this paper, several publications have also reported the analysis of antibody responses to N protein, S protein, and receptor-binding domain (RBD) on S protein in COVID-19 patients [14][15][16][17][18]. However, the seropositive rate of both IgM and IgG responses within one week after onset and in the context of both N protein and S protein has not been clarified.…”

Section: Introductionmentioning

confidence: 97%

Kinetics of SARS-CoV-2 specific IgM and IgG responses in COVID-19 patients

Sun

Feng

et al. 2020

Emerging Microbes & Infections

477

449

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The emerging COVID-19 caused by SARS-CoV-2 infection poses severe challenges to global public health. Serum antibody testing is becoming one of the critical methods for the diagnosis of COVID-19 patients. We investigated IgM and IgG responses against SARS-CoV-2 nucleocapsid (N) and spike (S) protein after symptom onset in the intensive care unit (ICU) and non-ICU patients. 130 blood samples from 38 COVID-19 patients were collected. The levels of IgM and IgG specific to N and S protein were detected by ELISA. A series of blood samples were collected along the disease course from the same patient, including 11 ICU patients and 27 non-ICU patients for longitudinal analysis. N and S specific IgM and IgG (N-IgM, N-IgG, S-IgM, S-IgG) in non-ICU patients increased after symptom onset. N-IgM and S-IgM in some non-ICU patients reached a peak in the second week, while N-IgG and S-IgG continued to increase in the third week. The combined detection of N and S specific IgM and IgG could identify up to 75% of SARS-CoV-2 infected patients in the first week. S-IgG was significantly higher in non-ICU patients than in ICU patients in the third week. In contrast, N-IgG was significantly higher in ICU patients than in non-ICU patients. The increase of S-IgG positively correlated with the decrease of C-reactive protein (CRP) in non-ICU patients. N and S specific IgM and IgG increased gradually after symptom onset and can be used for detection of SARS-CoV-2 infection. Analysis of the dynamics of S-IgG may help to predict prognosis.

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“…In the nucleocapsid, the viral genome is encapsulated by the nucleocapsid protein and thereby protected from the host cell environment (McBride et al, 2014;Gralinski and Menachery, 2020). The nucleocapsid protein of human coronaviruses is produced at high levels within infected cells and is critical for virion assembly (McBride et al, 2014;Fehr and Perlman, 2015;Gralinski and Menachery, 2020;Liu et al, 2020). In addition, it enhances the efficiency of sub-genomic viral RNA transcription and is essential for viral replication (McBride et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Nucleocapsid protein of SARS-CoV-2 phase separates into RNA-rich polymerase-containing condensates

Savastano

Opakua

Ranković

et al. 2020

Preprint

136

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The etiologic agent of the Covid-19 pandemic is the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The viral membrane of SARS-CoV-2 surrounds a helical nucleocapsid in which the viral genome is encapsulated by the nucleocapsid protein. The nucleocapsid protein of SARS-CoV-2 is produced at high levels within infected cells, enhances the efficiency of viral RNA transcription and is essential for viral replication. Here we show that RNA induces cooperative liquid-liquid phase separation of the SARS-CoV-2 nucleocapsid protein. In agreement with its ability to phase separate in vitro, we show that the protein associates in cells with stress granules, cytoplasmic RNA/protein granules that form through liquid-liquid phase separation and are modulated by viruses to maximize replication efficiency. Liquid-liquid phase separation generates high-density protein/RNA condensates that recruit the RNA-dependent RNA polymerase complex of SARS-CoV-2 providing a mechanism for efficient transcription of viral RNA. Inhibition of RNA-induced phase separation of the nucleocapsid protein by small molecules or biologics thus can interfere with a key step in the SARS-CoV-2 replication cycle.

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

Cited by 597 publications

References 17 publications

Sensitivity and specificity of a rapid test for assessment of exposure to SARS-CoV-2 in a community-based setting in Brazil

Sensitivity and specificity of a rapid test for assessment of exposure to SARS-CoV-2 in a community-based setting in Brazil

Kinetics of SARS-CoV-2 specific IgM and IgG responses in COVID-19 patients

Nucleocapsid protein of SARS-CoV-2 phase separates into RNA-rich polymerase-containing condensates

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