Journal: The Journal of Biological Chemistry

Article Title: Development of a highly specific and sensitive VHH-based sandwich immunoassay for the detection of the SARS-CoV-2 nucleoprotein

doi: 10.1016/j.jbc.2021.101290

Figure Lengend Snippet: Characterization of the Nucleoprotein . a shows the SDS Page gel with lanes 5 to 12 representing the eluted fractions containing the purified SARS-CoV-2 N and lane 13 to 18 are separated contaminants; b represents the intact mass measurement. The measured molecular weight (48 752.80 +/- 1.96 Da) is consistent with the expected average mass calculated from the full-length SARS-CoV-2 N primary sequence (48 752.13 Da, Δm = +0.67 Da (+13.7 ppm)) thereby confirming the structural integrity of the protein; c shows one main homogeneous population by DLS with a hydrodynamic radius of 6 nm. No aggregates are detectable at 37°C; d represents the AUC measurement where 96% of the sample is under a dimeric form.

Article Snippet: The Rapid SARS-CoV-2 Antigen Test Card (MP biomedicals) was used for the detection of N according to manufacturer’s instructions.

Techniques: SDS Page, Purification, Mass Measurement, Molecular Weight, Sequencing

Journal: The Journal of Biological Chemistry

Article Title: Development of a highly specific and sensitive VHH-based sandwich immunoassay for the detection of the SARS-CoV-2 nucleoprotein

doi: 10.1016/j.jbc.2021.101290

Figure Lengend Snippet: Analysis of the binding of the different VHHs to the SARS-CoV-2 Nucleoprotein . A) Binding of the different VHHs to SARS-CoV-2 recombinant Nucleoprotein determined by ELISA. N was coated at 1 μg/ml and VHHs at different concentrations were then added. B) Real-time monitoring of the VHH/N interaction by SPR. The determined kinetic parameters of the VHHS are provided in . C) Binding of the VHHs by ELISA on cell extracts. The VHH concentration leading to maximal difference obtained between infected and uninfected cell extracts are indicated by grey variations.

Article Snippet: The Rapid SARS-CoV-2 Antigen Test Card (MP biomedicals) was used for the detection of N according to manufacturer’s instructions.

Techniques: Binding Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Concentration Assay, Infection

Journal: The Journal of Biological Chemistry

Article Title: Development of a highly specific and sensitive VHH-based sandwich immunoassay for the detection of the SARS-CoV-2 nucleoprotein

doi: 10.1016/j.jbc.2021.101290

Figure Lengend Snippet: | Identification of the VHH binding sites by HDX-MS . a) General organization of full-length SARS-CoV-2 N showing the position of the two folded NTD and CTD structural domains and the three intrinsically disordered regions (N-arm, LKR, and C-tail). b) Differential fractional uptake plots showing the relative variations in deuterium incorporation imposed by the binding of each VHH to full-length SARS-CoV-2 N. The differences in uptake between the apo- and VHH-bound states were calculated for each peptide and time point and plotted as a function of peptide position. A positive uptake difference is indicative of a VHH-induced protective effect on the exchangeable amide hydrogens. Peptides displaying statistically significant uptake differences (Wald test, p< 0.01) are highlighted in gray (Figure S5). Peptides 340-353 (panel VHH E10-3), 337-353 (panel VHH E7-2) and 111-156 (panel VHH E4-3) were removed from the statistical analysis due to either poor fitting quality to the Mixed Effect Model or poor MS signal.

Article Snippet: The Rapid SARS-CoV-2 Antigen Test Card (MP biomedicals) was used for the detection of N according to manufacturer’s instructions.

Techniques: Binding Assay

Journal: The Journal of Biological Chemistry

Article Title: Development of a highly specific and sensitive VHH-based sandwich immunoassay for the detection of the SARS-CoV-2 nucleoprotein

doi: 10.1016/j.jbc.2021.101290

Figure Lengend Snippet: | Comparison of the VHH epitopes mapped by HDX-MS . a) Linear representation ( Left ) of full-length SARS-CoV-2 N with NTD in gray and CTD in light blue. The cartoon representations of the NTD domain (pdb # 7CDZ) and the CTD dimer (pdb # 7CE0) are shown on the right panel, with one CTD monomer colored light blue and the other colored black. The RNA-binding residues identified by NMR are also reported on the linear ( Left , blue bars) and the cartoon representations of the NTD domain ( Right , blue spheres). b,c) Comparison of the epitopes identified in the CTD (b) and the NTD (c) domains by HDX-MS. Red and orange patches correspond to regions were major and minor reductions in solvent accessibility were observed upon VHH binding. HDX-MS results are mapped onto the cartoon and surface representations of the NTD and CTD domains.

Article Snippet: The Rapid SARS-CoV-2 Antigen Test Card (MP biomedicals) was used for the detection of N according to manufacturer’s instructions.

Techniques: RNA Binding Assay, Binding Assay

Journal: The Journal of Biological Chemistry

Article Title: Development of a highly specific and sensitive VHH-based sandwich immunoassay for the detection of the SARS-CoV-2 nucleoprotein

doi: 10.1016/j.jbc.2021.101290

Figure Lengend Snippet: Immunofluorescence labeling of SARS-CoV-2 virus in infected cells . Representative staining with biotinylated VHHs at 1ug/mL of subconfluent layer of FRhK4 infected cells. A rabbit antibody against the SARS-CoV-2 N was used as a control of the infection.

Article Snippet: The Rapid SARS-CoV-2 Antigen Test Card (MP biomedicals) was used for the detection of N according to manufacturer’s instructions.

Techniques: Immunofluorescence, Labeling, Infection, Staining

Journal: The Journal of Biological Chemistry

Article Title: Development of a highly specific and sensitive VHH-based sandwich immunoassay for the detection of the SARS-CoV-2 nucleoprotein

doi: 10.1016/j.jbc.2021.101290

Figure Lengend Snippet: Immunofluorescence labeling of SARS-CoV-2 virus in the lung of infected syrian Hamster. Representative staining of lung slices with biotinylated VHHs at 1/500.Scales bar: 50 μm. left panel uninfected control, right panel infected hamster

Article Snippet: The Rapid SARS-CoV-2 Antigen Test Card (MP biomedicals) was used for the detection of N according to manufacturer’s instructions.

Techniques: Immunofluorescence, Labeling, Infection, Staining

Journal: The Journal of Biological Chemistry

Article Title: Development of a highly specific and sensitive VHH-based sandwich immunoassay for the detection of the SARS-CoV-2 nucleoprotein

doi: 10.1016/j.jbc.2021.101290

Figure Lengend Snippet: A: Immunofluorescence labeling of N protein in lung sections of mice infected with the B.1-351 and P1 SARS-Cov-2 variants .Representative staining of lung slices with biotinylated VHHs at 1/500.Scales bar: 50 μm. left panel uninfected control, right panels infected mice. B: Detection of Nucleoprotein from variants by sandwich ELISA. VHH NTD-E4-3 was coated on the plate, permeabilized SARS-CoV-2 virus variants were then added at different concentrations and were revealed by adding a biotinylated VHH G9-1 followed by peroxydase labelled streptavidin. Control without virus were performed and their values were substracted from the data.

Article Snippet: The Rapid SARS-CoV-2 Antigen Test Card (MP biomedicals) was used for the detection of N according to manufacturer’s instructions.

Techniques: Immunofluorescence, Labeling, Infection, Staining, Sandwich ELISA

Journal: PLoS ONE

Article Title: Field evaluation of the performance of seven Antigen Rapid diagnostic tests for the diagnosis of SARs-CoV-2 virus infection in Uganda

doi: 10.1371/journal.pone.0265334

Figure Lengend Snippet: Demographic characteristics of participants.

Article Snippet: There were 16 (6.6%) specimens that were MEDsan ® SARS-CoV-2 Antigen Rapid test positive ( ).

Techniques:

Journal: PLoS ONE

Article Title: Field evaluation of the performance of seven Antigen Rapid diagnostic tests for the diagnosis of SARs-CoV-2 virus infection in Uganda

doi: 10.1371/journal.pone.0265334

Figure Lengend Snippet: Field performance of 7 antigen RDTs for the diagnosis of SARs-CoV-2 virus in Uganda.

Article Snippet: There were 16 (6.6%) specimens that were MEDsan ® SARS-CoV-2 Antigen Rapid test positive ( ).

Techniques: Biomarker Discovery, Virus

Journal: PLoS ONE

Article Title: Field evaluation of the performance of seven Antigen Rapid diagnostic tests for the diagnosis of SARs-CoV-2 virus infection in Uganda

doi: 10.1371/journal.pone.0265334

Figure Lengend Snippet: COVID-19 Antigen Rapid test results compared to the Ct values on RT-PCR.

Article Snippet: There were 16 (6.6%) specimens that were MEDsan ® SARS-CoV-2 Antigen Rapid test positive ( ).

Techniques:

Journal: Micromachines

Article Title: Microfluidics-Based POCT for SARS-CoV-2 Diagnostics

doi: 10.3390/mi13081238

Figure Lengend Snippet: Some Commercialized POCT devices based on immunoassay.

Article Snippet: ANTIGEN RAPID TEST CASSETTE SARS-CoV-2 (SWAB) , A. Menarini Diagnostics, Italy , Immunoassay , 15 , Nasopharyngeal swab , 98.74% (95%CI: 96.80–99.66%) , 96.72% (95%CI: 88.65–99.60%) , 99.22% (95%CI: 97.21–99.91%) , Nucleocapside protein , [ ] .

Techniques: Sampling, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Diagnostic Assay

Journal: Eurosurveillance

Article Title: Can variants, reinfection, symptoms and test types affect COVID-19 diagnostic performance? A large-scale retrospective study of AG-RDTs during circulation of Delta and Omicron variants, Czechia, December 2021 to February 2022

doi: 10.2807/1560-7917.ES.2023.28.38.2200938

Figure Lengend Snippet: Sensitivities of SARS-CoV-2 AG-RDTs conducted on the same day as the PCR (suspected higher viral load cases), Information System of Infectious Diseases, Czechia, December 2021–February 2022 (total PCR-positive n = 5,488 for Delta and n = 18,966 for Omicron)

Article Snippet: VivaDiag Wellion SARS-CoV-2 Antigen Rapid Test - VivaChek Biotech (Hangzhou) Co., Ltd. , 1.6 , 2.3 , 83.3 , 86.9.

Techniques:

Journal: Eurosurveillance

Article Title: Role of population and test characteristics in antigen-based SARS-CoV-2 diagnosis, Czechia, August to November 2021

doi: 10.2807/1560-7917.ES.2022.27.33.2200070

Figure Lengend Snippet: Evaluation of SARS-CoV-2 antigen rapid diagnostic tests, Czechia, 5 August–6 December 2021 (n = 38 tests, n = 272,000 samples)

Article Snippet: 26 , (SARS-CoV-2) Antigen Rapid Test COVIDENT COVID-19 , Pierenkemper GmbH , 684 , 193 , 49.7 (42.2–57.3) , 96.1 (94.2–98.0).

Techniques: Diagnostic Assay