Journal: PLOS Neglected Tropical Diseases

Article Title: Engineered Nanobodies for early and accurate diagnosis of dengue virus infection

doi: 10.1371/journal.pntd.0013168

Figure Lengend Snippet: A-Various blocking agents and two distinct ELISA microplates were evaluated. The P/N ratio for NS1-2 capture indicated that 1% BSA, combined with Greiner microplates, yielded the highest P/N ratio. B-The ability of the Nbs under study to capture the NS1-2 protein was assessed using two different concentrations (10 and 20 μg/mL). Detection was performed using an anti-Flag HRP antibody. Nanobody 14 and Nb48 exhibited a stronger capture capacity compared to other tested Nbs.

Article Snippet: Although there are several commercial DENV NS1 ELISA kits available, in Argentina, but also in Latin-American countries, most public hospitals use Platelia Dengue NS1 Ag kit (BioRad) and Panbio Dengue Early ELISA kit (Abbott).

Techniques: Blocking Assay, Enzyme-linked Immunosorbent Assay

Journal: PLOS Neglected Tropical Diseases

Article Title: Engineered Nanobodies for early and accurate diagnosis of dengue virus infection

doi: 10.1371/journal.pntd.0013168

Figure Lengend Snippet: A-Schematic representation of the two proposed Nanobody-based ELISA systems. System 1: 4 µg/mL of Nb14 Fc as the capture antibody and 10 μg/mL Nb48 HRP as the detection antibody. System 2: 2 µg/mL of Nb14 Fc as the capture antibody and 5 μg/mL biotinylated Nb66 as the detection antibody. B-Reactivity of System 1 with NS1 variants from DENV serotypes 1–4, ZIKV NS1, and human sera. C-Reactivity of System 2 with the same antigen panel. Data is shown as mean absorbance at 450 nm ± SD (n=3). Black dashed line represents the mean OD of negative control samples plus 3 standard deviations.

Article Snippet: Although there are several commercial DENV NS1 ELISA kits available, in Argentina, but also in Latin-American countries, most public hospitals use Platelia Dengue NS1 Ag kit (BioRad) and Panbio Dengue Early ELISA kit (Abbott).

Techniques: Enzyme-linked Immunosorbent Assay, Negative Control

Journal: PLOS Neglected Tropical Diseases

Article Title: Engineered Nanobodies for early and accurate diagnosis of dengue virus infection

doi: 10.1371/journal.pntd.0013168

Figure Lengend Snippet: A-Schematic representation of the combined Nanobody ELISA workflow. B-Detection of recombinant NS1 proteins from DENV and ZIKV in a negative serum matrix. C-Detection of NS1 in pooled human serum samples from DENV-positive and negative patients. Serum dilutions (1:5 and 1:10) were analyzed using biotinylated Nb66 as detector. D-Specificity assessment of the combined Nb ELISA using supernatants from Vero cells infected with different flaviviruses (DENV-1 to DENV-4, ZIKV, YFV) and the unrelated arbovirus CHIKV, which co-circulate in endemic regions and cause overlapping clinical symptoms. Mock-infected supernatant was used as a negative control.

Article Snippet: Although there are several commercial DENV NS1 ELISA kits available, in Argentina, but also in Latin-American countries, most public hospitals use Platelia Dengue NS1 Ag kit (BioRad) and Panbio Dengue Early ELISA kit (Abbott).

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Infection, Negative Control

Journal: PLOS Neglected Tropical Diseases

Article Title: Engineered Nanobodies for early and accurate diagnosis of dengue virus infection

doi: 10.1371/journal.pntd.0013168

Figure Lengend Snippet: A-ELISA to detect DENV1-4 NS1 at different concentrations to determine the limit of detection of the optimized cNb-ELISA. The developed detection system has variable LOD for the different DENV NS1. B-Estimation of the analytical sensitivity of the cNb-ELISA, absorbance values obtained based on 2-fold serial dilutions of the pool of positive sera in a range from 1/2 to 1/256. Inset: linear regression analysis of absorbance values based on antigen concentration.

Article Snippet: Although there are several commercial DENV NS1 ELISA kits available, in Argentina, but also in Latin-American countries, most public hospitals use Platelia Dengue NS1 Ag kit (BioRad) and Panbio Dengue Early ELISA kit (Abbott).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Journal of Medical Virology

Article Title: Seroprevalence and Placental Transfer of Zika and Dengue Virus Antibodies in Postpartum Women in Southeast Brazil

doi: 10.1002/jmv.70384

Figure Lengend Snippet: Flowchart of the serological tests conducted to detect recent or past ZIKV and DENV infections in serum samples of all postpartum women participants ( n = 601). The samples were screened using an ELISA to detect specific IgG antibodies with all positive results confirmed by the CPE‐VNT technique to detect neutralizing antibodies. Discordant samples were further analyzed using the PRNT assay. Once true positive samples were identified they were subsequently tested for the presence of anti‐ZIKV and anti‐DENV specific IgM antibodies.

Article Snippet: For the detection of anti‐ZIKV and anti‐DENV IgM antibodies, the anti‐Zika virus IgM μ‐capture ELISA kit (#ab213327, Abcam, Cambridge, USA) and the Panbio Dengue IgM Capture ELISA kit (#01PE20, Abbott, Chicago, IL, USA) were used, respectively, following the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Plaque Reduction Neutralization Test