Journal: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America

Article Title: Distinguishing Secondary Dengue Virus Infection From Zika Virus Infection With Previous Dengue by a Combination of 3 Simple Serological Tests

doi: 10.1093/cid/cix672

Figure Lengend Snippet: Results of Enzyme-Linked Immunosorbent Assays in Different Serum/Plasma Panels

Article Snippet: Purified DENV1-NS1 protein was purchased from the Native Antigen (United Kingdom).

Techniques:

Journal: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America

Article Title: Distinguishing Secondary Dengue Virus Infection From Zika Virus Infection With Previous Dengue by a Combination of 3 Simple Serological Tests

doi: 10.1093/cid/cix672

Figure Lengend Snippet: Sensitivity and Specificity of Enzyme-Linked Immunosorbent Assays

Article Snippet: Purified DENV1-NS1 protein was purchased from the Native Antigen (United Kingdom).

Techniques:

Journal: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America

Article Title: Distinguishing Secondary Dengue Virus Infection From Zika Virus Infection With Previous Dengue by a Combination of 3 Simple Serological Tests

doi: 10.1093/cid/cix672

Figure Lengend Snippet: Results of nonstructural protein 1 (NS1) immunoglobulin G (IgG) enzyme-linked immunosorbent assays (ELISAs) for secondary dengue virus (sDENV) and Zika virus (ZIKV) with previous (wp) DENV infection panels. DENV1-NS1 (A) and ZIKV-NS1 (B) IgG ELISAs in convalescent-phase samples from sDENV, ZIKVwpDENV (Nicaragua [Nic]), probable ZIKVwpDENV (Brazil [Bra]) and negative control (NC) panels. C, Relative optical density (rOD) ratio of ZIKV-NS1 to DENV1-NS1. The sensitivity and specificity are shown based on a cutoff rOD ratio at 0.24. DENV1-NS1 (D) and ZIKV-NS1 (E) IgG ELISAs in post–convalescent phase samples (3 months to 2 years post–symptom onset) from sDENV panels. F, Positive rates of DENV1- and ZIKV-NS1 IgG ELISAs in sDENV panels over time. Dotted lines indicate cutoff rOD values for ELISAs. Data are mean of 2 experiments (each in duplicate). Two-tailed Mann-Whitney test was used to compare the 2 groups.

Article Snippet: Purified DENV1-NS1 protein was purchased from the Native Antigen (United Kingdom).

Techniques: Virus, Infection, Negative Control, Two Tailed Test, MANN-WHITNEY

Journal: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America

Article Title: Distinguishing Secondary Dengue Virus Infection From Zika Virus Infection With Previous Dengue by a Combination of 3 Simple Serological Tests

doi: 10.1093/cid/cix672

Figure Lengend Snippet: Results of nonstructural protein 1 (NS1) immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISAs) in sequential samples from individuals with secondary dengue virus (sDENV) and Zika virus (ZIKV) infection with previous (wp) DENV infections. A, Three cases with sDENV infection, 3–18 months post–symptom onset. B, Four blood donors with sDENV infection, 7 days to 6 months post–index day (PID). C, Five blood donors with ZIKVwpDENV infection, from index day to 3 months PID. Three cases (1, 4, and 7) seroconverted to ZIKV-NS1; 2 cases (15 and 21) had ZIKV-NS1 IgG starting from the index day. Dotted lines indicate cutoff relative optical density (rOD) values for ELISAs. Data are mean of 2 experiments (each in duplicates).

Article Snippet: Purified DENV1-NS1 protein was purchased from the Native Antigen (United Kingdom).

Techniques: Enzyme-linked Immunosorbent Assay, Virus, Infection

Journal: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America

Article Title: Distinguishing Secondary Dengue Virus Infection From Zika Virus Infection With Previous Dengue by a Combination of 3 Simple Serological Tests

doi: 10.1093/cid/cix672

Figure Lengend Snippet: Proposed algorithm of using 3 serological tests (without neutralization tests) to distinguish different Zika virus (ZIKV) and dengue virus (DENV) infections in dengue- and Zika-endemic regions in the framework of Centers for Disease Control and Prevention guidelines for laboratory diagnosis of ZIKV infection [6, 7]. Only samples tested positive or equivocal by ZIKV or DENV E protein–based immunoglobulin M (IgM) enzyme-linked immunosorbent assays (ELISAs) are included in the algorithm. The total numbers from each panel and the numbers of positive or negative based on the 3 Nonstructural Protein 1 (NS1) ELISAs are shown in parentheses. Abbreviations: IgG, immunoglobulin G; OD, optical density; pDENV, primary dengue virus infection; pZIKV, primary Zika virus infection; rOD, relative OD; sDENV, secondary dengue virus infection; ZIKVwpDENV, Zika virus infection with previous dengue virus infection.

Article Snippet: Purified DENV1-NS1 protein was purchased from the Native Antigen (United Kingdom).

Techniques: Neutralization, Virus, Infection

Journal: Plant Biotechnology Journal

Article Title: Plant‐made dengue virus‐like particles produced by co‐expression of structural and non‐structural proteins induce a humoral immune response in mice

doi: 10.1111/pbi.13501

Figure Lengend Snippet: Analysed candidate vaccines and Coomassie‐stained SDS‐PAGE control: The respective vaccine candidates were expressed in plants, purified and analysed by SDS‐PAGE. DENV1‐VLPs were produced by co‐expressing DENV1‐SP and NSP. SDS‐PAGE shows signals for prM (18 kDa) and E (~55 kDa). Empty bluetongue core‐like particles (BTV‐CLPs) as negative control (c‐) were produced by co‐expression of BTV VP3 (103 kDa) and VP7 (39 kDa). For antigen display inside BTV‐CLPs, the E domain III (EIII) of DENV1, DENV4 and ZIKA was fused to the N terminus of BTV‐VP3. Co‐expression of BTV‐VP3 and V7 leads to assembly of CLPs and integration of the N ‐terminal fused antigens. The Coomassie‐stained SDS‐PAGE shows the size difference between c‐ VP3 and VP3 fused to EIII. The EIII soluble subunit (14 kDa) showed the expected size of 14 kDa and high purity.

Article Snippet: To detect specific antibodies, 96‐well plates (NUNC Maxisorp) were coated with 5 µg/mL of each of the following antigens diluted in carbonate–bicarbonate buffer (Thermo, Product #28382): recombinant Zika virus envelope protein (Native Antigen Company, ZIKVSU‐ENV‐100), Zika VLP (Native Antigen Company, ZIKV‐VLP‐250), DENV1 VLP (Native Antigen Company, DENV1‐VLP‐250) and DENV4 VLP (Native Antigen Company, DENV4‐VLP‐250).

Techniques: Vaccines, Staining, SDS Page, Purification, Produced, Expressing, Negative Control

Journal: Plant Biotechnology Journal

Article Title: Plant‐made dengue virus‐like particles produced by co‐expression of structural and non‐structural proteins induce a humoral immune response in mice

doi: 10.1111/pbi.13501

Figure Lengend Snippet: Humoral response of DENV1 VLPs, EIII domains displayed in bluetongue CLPs and the DENV1‐EIII subunit. Six mice per group were immunized in a single or prime‐boost manner. Collected sera of immunized mice were tested in a dilution series against commercial DENV1, DENV4 and ZIKA VLPs, as well as recombinant ZIKA envelope protein (ZIKV‐Env). Lines show mean values with error bars denoted standard error. Only DENV1‐VLPs stimulated the production of reacting antibodies, indicating that in mice, VLPs are more efficient antigens in terms of B‐cell response than comparable amounts of the DENV1 subunit alone or the display of EIII on bluetongue CLPs.

Article Snippet: To detect specific antibodies, 96‐well plates (NUNC Maxisorp) were coated with 5 µg/mL of each of the following antigens diluted in carbonate–bicarbonate buffer (Thermo, Product #28382): recombinant Zika virus envelope protein (Native Antigen Company, ZIKVSU‐ENV‐100), Zika VLP (Native Antigen Company, ZIKV‐VLP‐250), DENV1 VLP (Native Antigen Company, DENV1‐VLP‐250) and DENV4 VLP (Native Antigen Company, DENV4‐VLP‐250).

Techniques: Recombinant