Journal: Frontiers in Microbiology

Article Title: A Peptide-Based Virus Inactivator Protects Male Mice Against Zika Virus-Induced Damage of Testicular Tissue

doi: 10.3389/fmicb.2019.02250

Figure Lengend Snippet: Z2 inhibited infection of ZIKV strains with Asian and African lineages in TM4 cells. (A) Sequence and location of Z2 in ZIKV E dimer. Side view of ZIKV E protein in the dimeric conformation is shown [Protein Data Bank: 5IRE ]; DI, DII, DIII, stem and transmembrane domain are shown in red, yellow, blue, orange, and wheat, respectively; the fusion peptide is shown in cyan, and Z2 (421–453) is shown in green. TM4 cells were infected with ZIKV strain SZ01 (B) , MR766 (C) and FLR (D) , treated with Z2 at different concentration, and then viral copies in the supernatant at 48 h were measured by qRT-PCR. Data were presented as means ± SD. (E) Z2-treated ZIKV of different strain lost infectivity on TM4 cells irreversibly. After incubation with Z2 at 37°C for 2 h, ZIKV particles were separated from the unbound Z2 by PEG 8000 to measure their infectivity on TM4 cells. Each sample was tested in triplicate and the experiments were repeated at least once. The data from two independent experiments were presented as mean ± SD.

Article Snippet: ZIKV strains MR766 (#VR1838) and FLR (#VR1844) were obtained from ATCC.

Techniques: Infection, Sequencing, Concentration Assay, Quantitative RT-PCR, Incubation

Journal: Science translational medicine

Article Title: Rapid Antigen Tests for Dengue Virus Serotypes and Zika Virus in Patient Serum

doi: 10.1126/scitranslmed.aan1589

Figure Lengend Snippet: (A) Table listing mAb names, mAb immunochromatography applications, mAb linear epitope sequences and starting amino acid positions, and NS1 domain positions. (B) Comparison of amino acid similarity based on analysis of NS1 protein sequences from the following viruses: DENV1- Strain Singapore/S275/1990, accession number P33478; DENV2 -Strain NGC, accession number AAA42941; DENV3- Philippines/H87/1956, accession number AAA99437; DENV4- Singapore/8976/1995, accession number AAV31422; Zika virus, accession number KU497555.1. Amino acid sequences were compared using Color Align Conservation http://www.bioinformatics.org/sms2/color_align_cons.html to enhance the output of sequence alignment program. Residues that are identical among the sequences are boxed. Linear peptide epitopes (B) are italicized and indicated in color on the figure, with the key to the right of the figure.

Article Snippet: BALB-c mice were immunized with purified recombinant DENV or ZIKV NS1 proteins that were expressed in mammalian cells (Native Antigen Co.) to facilitate protein folding and post-translational modifications.

Techniques: Comparison, Virus, Sequencing

Journal: Science translational medicine

Article Title: Rapid Antigen Tests for Dengue Virus Serotypes and Zika Virus in Patient Serum

doi: 10.1126/scitranslmed.aan1589

Figure Lengend Snippet: (A-B) Images of rapid test strips. Strip numbers refer to the DENV serotype NS1 (1–4), pan-dengue (P; all four DENV serotype NS1 proteins), or ZIKV virus NS1 (Z) detected. Recombinant NS1 proteins, indicated with a lower case “r’ preceding the virus name, were prepared at 500 ng/ml, and the strips were run using 50 μl of solution. Strip #1 (detects DENV serotype 1): mAb pair 912/271; Strip #2 (detects DENV serotype 2): mAb pair 243/1; Strip #3 (detects DENV serotype 3): mAb pair 411/55; Strip #4 (detects DENV serotype 4): mAb pair 626/55; Strip P (“pan dengue”; detects all four DENV serotypes): mAbs 271–243-411–626/323; Strip Z (detects ZIKV): mAbs 130/110. The test proteins run on the strips are recombinant DENV NS1, serotypes 1–4 (rDENV1-rDENV4), as well as recombinant NS1 proteins from ZIKV (rZIKV), West Nile Virus (rWNV), Yellow Fever Virus (rYFV), Japanese encephalitis virus (rJEV), and Tick Borne encephalitis virus (rTBEV). C = control, NS1 = detection site for specific NS1 protein. (C-E) Limits of detection for viral NS1 proteins using the serotype-specific (SSp) DENV strips 1–4 (C), the pan-dengue strip (D), and the ZIKV virus strip (E). The limits of detection, representing three independent determinations, are recorded on the figures. Each point (C-E) is presented as the mean and standard deviation. (F) NS1-containing supernatants from Vero cells infected with DENV serotypes 1–4 (Vs DENV1–4) or ZIKV virus (Vs ZIKV-U (Uganda) or ZIKV-B (Brazil)) were chromatographed on strips 1–4, pan-dengue (P), or ZIKV (Z) NS1 strips. The arrows indicate the strips with positive NS1 signals. Horizontal test lines (panel F) result from applying antibodies to the nitrocellulose using a mechanical striper device; the circular dot signals result from applying antibodies to the nitrocellulose using a standard pipettor.

Article Snippet: BALB-c mice were immunized with purified recombinant DENV or ZIKV NS1 proteins that were expressed in mammalian cells (Native Antigen Co.) to facilitate protein folding and post-translational modifications.

Techniques: Stripping Membranes, Virus, Recombinant, Standard Deviation, Infection

Journal: Science translational medicine

Article Title: Rapid Antigen Tests for Dengue Virus Serotypes and Zika Virus in Patient Serum

doi: 10.1126/scitranslmed.aan1589

Figure Lengend Snippet: (A) Map showing the endemic virus regions where the rapid tests were deployed to analyze patient serum samples. The areas of the circles correlate with the numbers of samples analyzed. The blue colors, faint to dark, represent DENV serotypes 1–4. ZIKV is indicated in orange color. (B) ELISA results showing the amounts of DENV (left) and ZIKV (right) NS1 found in patient serum and supernatants from infected cell cultures. Lanes 1 and 6 are supernatants from Vero cells infected with DENV; lanes 2 and 7 are supernatants from Vero cells infected with ZIKV. Lanes 3 and 8 are PCR-negative sera; lanes 4 and 9 are sera from PCR-positive DENV patients. Lanes 5 and 10 are sera from PCR-positive ZIKV patients. (C) Images of rapid test analysis of DENV NS1 serotypes 1–4 and ZIKV NS1 on serotype specific strips 1–4, as well as pan-dengue (P) and ZIKV (Z); the upward arrows mark positive tests, and θ is serum from an uninfected patient. (D-G) Quantification of rapid test results. Dipstick tests were run with PCR-confirmed DENV sera or ELISA-validated ZIKV serum (panel C), and the resulting signals were quantified and expressed as box plots. Statistical significance, based on one one-way ANOVA, is indicated as * P < 0.05, ** P < 0.01, *** P < 0.001. (H) Statistical significance, based on an unpaired T-test, is presented as * P < 0.05. Box and whiskers plots: the black ✕ represent the maximum and minimum measured normalized intensity values, while the small square box ☐ represents the mean value, and the larger box represents the 25–75% range of the data. Individual colored points represented individual patient samples measured. (I-J) Images of rapid tests showing that DENV and ZIKV NS1 tests do not cross-react. (I) Supernatants from Vero cells infected with DENV serotype 4 were chromatographed on DENV serotype strips 1–4, on the pan-dengue strip (P), and on the ZIKV NS1 strip (Z). (J) Supernatants from Vero cells infected with ZIKV virus were chromatographed on DENV serotype strips 1–4, on the pan-dengue strip (P), and on the ZIKV NS1 strip (Z). (K) Images of rapid tests showing ZIKV NS1 is detected in serum samples concentrated 5X, but ZIKV virus NS1 is not detected in concentrated urine. Three sets of paired serum and urine samples were concentrated 5X by filter centrifugation and chromatographed on the ZIKV dipsticks. S: serum; U: urine. (I-K) The red boxes and vertical black lines serve as fiducial markers for image recognition and processing. Upward arrows indicate positive tests, using the serum samples. (L-N) Quantification of NS1 protein in supernatants of Vero cells infected separately with three DENV4 patient isolates (L) or three ZIKV patient isolates (M), or five paired serum/urine patient samples (N). Fig. 3L-N: One-way ANOVA was used to calculate statistical significance of the dengue and Zika tests: p<0.05, p<0.01, and p<.0.001 are indicated as *, **, and ***, respectively. Box and whiskers plots: the black ✕ represent the maximum and minimum measured normalized intensity values, while the black ☐ represents the mean value, and the larger box represents the 25–75% range of the data. Individual colored points represented individual patient samples measured.

Article Snippet: BALB-c mice were immunized with purified recombinant DENV or ZIKV NS1 proteins that were expressed in mammalian cells (Native Antigen Co.) to facilitate protein folding and post-translational modifications.

Techniques: Virus, Enzyme-linked Immunosorbent Assay, Infection, Stripping Membranes, Centrifugation

Journal: Viruses

Article Title: Evaluation of the LIAISON XL Zika Capture IgM II for the Diagnosis of Zika Virus Infections

doi: 10.3390/v12010069

Figure Lengend Snippet: Final resolution of samples with discrepant results.

Article Snippet: Finally, the ZIKV plaque reduction neutralization test (PRNT) was carried out with an in-house method using Vero E6 cells (ATCC CRL-1586, American Type Cell Collection, Manassas, Virginia) and 100 TCID50 of ZIKV (African strain MR-766, isolated from a rhesus monkey in the Zika Forest, Uganda, in 1947, and maintained in Vero E6 cells).

Techniques:

Journal: Applied Microbiology and Biotechnology

Article Title: Recent advances in the potential applications of luminescence-based, SPR-based, and carbon-based biosensors

doi: 10.1007/s00253-022-11901-6

Figure Lengend Snippet: Advances and potential application of biosensors in pharmaceutical sciences

Article Snippet: Electrochemical immunosensor , Zika virus (envelope protein) , 10 pM , Zika virus (strain Zika SPH2015) envelope protein (ZIKV-E) ELISA Pair Set; Sino Biological Inc. (LOD = 125 pg/mL) , Promising clinical application for early-stage diagnostics of the virus, operation time around 40 min , Kaushik et al. ( ) .

Techniques: Förster Resonance Energy Transfer, Imaging, SPR Assay, Electrochemiluminescence, MicroChIP Assay, In Vitro, Infection, Amplification, Isolation, Biomarker Assay, Sequencing

Journal: Applied Microbiology and Biotechnology

Article Title: Recent advances in the potential applications of luminescence-based, SPR-based, and carbon-based biosensors

doi: 10.1007/s00253-022-11901-6

Figure Lengend Snippet: Summarization of LOD, advances of the abovementioned biosensors, and their comparison with common methods of detection

Article Snippet: Electrochemical immunosensor , Zika virus (envelope protein) , 10 pM , Zika virus (strain Zika SPH2015) envelope protein (ZIKV-E) ELISA Pair Set; Sino Biological Inc. (LOD = 125 pg/mL) , Promising clinical application for early-stage diagnostics of the virus, operation time around 40 min , Kaushik et al. ( ) .

Techniques: Glutamate Assay, Enzyme-linked Immunosorbent Assay, Diagnostic Assay, Sequencing, Simultaneous Assay, Biomarker Assay, Fluorescence, In Situ Hybridization