Journal: bioRxiv

Article Title: Modeling herpes simplex virus type 2 and Zika virus replication in vaginal organoids and spheroids

doi: 10.64898/2026.03.02.709097

Figure Lengend Snippet: NS1 concentration in the supernatant (A) and NS5 transcript expression (B) in mouse vaginal organoids infected with ZIKV at MOI of 1. Values normalized to uninfected samples (Ni) and reported as fold change. (C) NS4B staining in organoids sections from 48 hours post-infection. ZIKV titer of mouse vaginal organoids (D) and VK2 spheroids (E) infected with ZIKV in the presence of 10 µM RDV. Values are mean ± SEM of 3 biological replicates. One-way ANOVA or two-way ANOVA with Tukey’s test for multiple comparisons was performed for statistical analysis. *, p < 0.05; **, p < 0.01; ***, p < 0.001. Magnification 63X; scale bars represent 50 µm. LD, limit of detection (5 PFU/ml). h, hours post-infection.

Article Snippet: ZIKV titer from infection experiments was quantified using an ELISA to detect NS1 concentration (Sino Biological, Cat No. KIT40544).

Techniques: Concentration Assay, Expressing, Infection, Staining

Journal: bioRxiv

Article Title: Dengue NS1 Antibodies drive Immune Complex Formation, Hyperglycaemia and systemic pathology in a murine NS1 plasmid challenge model

doi: 10.1101/2025.09.25.678465

Figure Lengend Snippet: (A–B) DV2 infection kinetics in pancreatic β-cell line. (A) Min6 cells were infected with DV2_LS, and samples were collected at indicated time points. Viral genome equivalents (gE) were quantified in extracellular (EC, 1□mL supernatant) and intracellular (IC, 300□µL lysate) fractions using qRT-PCR. Trypan blue exclusion was used to assess cell viability. Column graphs represent the percentage of live cells; line graphs depict viral load over time. (B) Total secreted NS1 levels (ng/mL) at different time points in the supernatant of infected Min6 cells were measured via quantitative NS1 ELISA. (C–D) GAPDH protein expression following DV infection. (C) Mean fold change in GAPDH expression (normalized to β-tubulin) in DV-infected Huh7 cells (all serotypes combined) versus uninfected controls at 96 hpi. (D) Serotype-specific fold changes in GAPDH expression in Huh7 cells infected with individual DV serotypes at 96 hpi. (E–F) Effect of NS1 Ab on GAPDH expression in hepatic cells. (E) Mean fold change in GAPDH expression in Huh7 cells treated with NS1 Ab-positive sera compared to control mouse sera at 96 hours. (F) Fold changes in GAPDH expression following treatment of Huh7 cells with NS1 Ab-positive sera from different DV serotype-specific NS1 administered mice compared to control sera at 96 hours. Data represented as mean±SEM.

Article Snippet: Detection of DV NS1 Ab was performed by ELISA using Human Anti-Dengue Virus IgG ELISA kit according to the manufacturer’s protocol with few modifications (R&D Systems, Cat# DENG00, MN, USA).

Techniques: Infection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing, Control

Journal: PLOS One

Article Title: Neural and endothelial cell-derived extracellular vesicles mediate Zika virus genome dissemination and productive infection in vivo

doi: 10.1371/journal.pone.0337609

Figure Lengend Snippet: Infection of all three cell types was confirmed by IF detection of the ZIKV C protein (green) at 48 hpi. Infected cells were identified through co-staining with cell-type specific markers (red). While all cultures demonstrated susceptibility to ZIKV infection, (A) neurons exhibited the most pronounced morphological alteration compared to uninfected controls. Notably, viral antigen is localized not only to perinuclear region but also to the axon hillock (arrow) and neurites (arrowhead). In contrast, (B) astrocytes and (C) MBECs maintained their typical cellular architecture despite infection. The highlighted square denotes a region of interest shown at higher magnification. Nuclei were counterstained with DAPI (blue). Scale bars represent 20 µm for neurons and MBECs, and 50 µm for astrocytes. Images are representative of two independent experiments, each comprising three replicates and eight fields analyzed per slide.

Article Snippet: Following this period, some wells were fixed with 4% PFA and processed by immunoperoxidase staining, using the ZIKV capsid antibody (Novus, NBP3–13200).

Techniques: Infection, Staining

Journal: PLOS One

Article Title: Neural and endothelial cell-derived extracellular vesicles mediate Zika virus genome dissemination and productive infection in vivo

doi: 10.1371/journal.pone.0337609

Figure Lengend Snippet: Immunoperoxidase staining for ZIKV E protein in A549 cell inoculated for 72 h with EVs or ZIKV. (A) Controls included non-infected cells (NIC), mock-treated cells (supernatants from uninfected C6/36HT cells), and supernatants of ZIKV-infected cells at a MOI of 0.1. (B) Brown peroxidase signal indicated productive infection in cells exposed to EVs-IC, while no detectable staining was observed in EVs-GlyR-treated cells. Representative images from three independent experiments, each performed in triplicate. Scale bars: 100 µm for controls and EVs-GlyR) and 50 µm for EVs-IC.

Article Snippet: Following this period, some wells were fixed with 4% PFA and processed by immunoperoxidase staining, using the ZIKV capsid antibody (Novus, NBP3–13200).

Techniques: Immunoperoxidase Staining, Infection, Staining

Journal: Science Advances

Article Title: Virus detection using nanoparticles and deep neural network–enabled smartphone system

doi: 10.1126/sciadv.abd5354

Figure Lengend Snippet: Pt-nanoprobes preparation: ( A ) TEM image and particle size distribution histogram. ( B ) Schematic of Pt-nanoprobes structure and UV-vis absorption analysis showing spectra of PtNPs (citrate-capped PtNPs, black) and Pt-nanoprobes (PtNPs conjugated to mAb, red). Photo credit: Mohamed S. Draz, Brigham and Women’s Hospital. Virus capture and labeling on-chip: ( C ) SDS-PAGE analysis of ZIKV released from the surface of the chip. Lane M, protein marker; lane ZIKV on chip, ZIKV released from the chip; lane ZIKV, ZIKV standard solution (10 6 particles/ml, virus not added to the chip). Rectangles in red and blue edges mark bands from antibodies and ZIKV, respectively. ( D ) ELISA and RT-PCR assay results to detect and quantify the captured ZIKV on-chip. Error bars are SDs from two independent experiments. ( E ) SEM images of ZIKV captured on chip and labeled with Pt-nanoprobes: (i) ZIKV on chip and (ii) ZIKV captured and labeled with Pt-nanoprobes. The concentration of the virus sample was 10 6 particles/ml. The results are expressed as the average values of at least three independent experiments.

Article Snippet: The surface of the cleaned glass slides was activated using PE-25 oxygen plasma (100 mW, 15% oxygen; Plasma Etch Inc., Carson City, NV, USA) for 2 to 3 min and modified using silane-functionalized PEG thiol (Nanocs Inc., New York, NY, USA) and mAb for capture of the target virus: anti-ZIKV envelope antibody (catalog no. HM325, EastCoast Bio Inc., North Berwick, ME, USA) for ZIKV, anti-HBV surface antigen (Ad/Ay) antibody (catalog no. ab54247, Abcam, Cambridge, MA, USA) for HBV, and anti-HCV core antigen antibody (catalog no. ab2582, Abcam, Cambridge, MA, USA) for HCV.

Techniques: Virus, Labeling, SDS Page, Marker, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Concentration Assay

Journal: Cell

Article Title: Rapid, Low-Cost Detection of Zika Virus Using Programmable Biomolecular Components.

doi: 10.1016/j.cell.2016.04.059

Figure Lengend Snippet: Figure 5. NASBA-CRISPR Cleavage (NASBACC) Allows for Strain Differentiation at Single-Base Resolution (A) Schematic representation of NASBACC genotyping following a positive Zika diagnosis. A synthetic trigger sequence is appended to a NASBA-amplified RNA fragment through reverse transcription. The presence of a strain-specific PAM leads to the production of either truncated or full-length trigger RNA, which differentially activates a toehold switch (sensor H) (Pardee et al., 2014). (B) The probability that a non-biased single nucleotide polymorphism (SNP) between two strains can be discriminated by CRISPR/Cas9 is 48% (Table S4). Hence, genetic drift between the American and African or Asian strains, while relatively small (14.4% and 4.9% sequence dissimilarity, respectively), has created hundreds of strain-specific PAM sites. (C) A SNP between African (GenBank: KF268950) and American (GenBank: KU312312) strains at site 7330 disrupts an existing PAM site, allowing for Cas9- mediated DNA cleavage only in the American strain. (D) Sensor 32B can distinguish between Dengue and Zika RNA sequences but cannot discriminate between American and African Zika strains. Paper discs containing sensor 32B were rehydrated with 300 nM trigger RNA corresponding to sequences from American-Zika, African-Zika, or Dengue. Colorimetric outputs: a purple color indicates the activation of LacZ expression from the toehold switch, and a yellow color indicates the toehold switch remained inactive. (E) NASBACC can discriminate between American- and African-lineages of Zika virus. Paper discs containing sensor H were rehydrated with a 1:10 dilution of NASBACC reactions initiated with 0.05 ml of a 300 nM RNA sample. In this case, an inactive toehold switch leads to a positive identification of the American Zika strain.

Article Snippet: Plasmids for sensors 27B and 32B are available through Addgene (plasmid numbers: 75006–75011).

Techniques: CRISPR, Biomarker Discovery, Sequencing, Reverse Transcription, Activation Assay, Expressing, Virus