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Subject: search.filters.subject.Dengue virus

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    Laser-induced graphene electrochemical immunosensors for rapid and sensitive serological detection: A case study on dengue detection platform
    (2025-06-01)
    Inlumphan, Supawee
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    Wongwiriyapan, Winadda
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    Khemasiri, Narathon
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    Rattanawarinchai, Prapakorn
    ;
    Leepheng, Piyawan
    Here, we present a diagnostic on a chip platform based on laser-induced graphene (LIG) electrochemical immunosensors for serological detection. The electrochemical immunosensors are fabricated through CO<inf>2</inf> laser induction on polyimide (PI) sheets. Optimal electrochemical activity of LIG electrodes is obtained under optimized conditions of laser fluence. To verify the application, the serological detection platform was demonstrated. After functionalization with dengue virus (DENV) antigen, the LIG electrochemical immunosensors are able to sense the presence of mouse anti-flavivirus monoclonal (4G2) antibody in a wide linear working range of 25–20,000 ng/ml with the limit of detection (LOD) of 17.41 ng/ml. A specific recognition with 4G2 antibodies against with media protein and isotype is confirmed. Furthermore, the reliability of LIG electrochemical immunosensors compared to conventional enzyme-linked immunosorbent assay (ELISA) is verified through the NS1 antibodies identification in human blood serum clinical samples at room temperature. Our results highlight that the LIG-based electrode is a promising platform for electrochemical immunosensors, aimed at developing reliable and practical diagnostic tools for serological detection. These tools enable early diagnosis of infectious diseases, as well as non-invasive and rapid screening.
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    Single-cell RNA sequencing reveals the expansion of circulating tissue-homing B cell subsets in secondary acute dengue viral infection
    (2024-05-30)
    Arora, Jantarika Kumar
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    Matangkasombut, Ponpan
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    Charoensawan, Varodom
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    Opasawatchai, Anunya
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    Sakuntabhai, Anavaj
    The roles of antibodies secreted by subsets of B cells in dengue virus (DENV) infection have been extensively studied, yet, the contribution of tissue-homing B cells to antiviral immunity remains unclear. In this study, we performed a comprehensive analysis of B cell subpopulations in peripheral blood samples from DENV-infected patients using single-cell RNA-sequencing (scRNA-seq) datasets and flow cytometry. We showed that plasma cells (PCs) and plasmablasts (PBs) were the predominant B cell populations during the acute phase of secondary natural DENV infection, but not in convalescent phase nor in healthy controls. Interestingly, these cells expressed proliferation, adhesion, and tissue-homing genes, including SELPLG, a homing marker of the skin, the initial infected site of DENV. Flow cytometry analysis confirmed a significant upregulation of cell surface expression of a cutaneous lymphocyte-associated antigen (CLA) encoded by SELPLG in PCs and PBs, compared to naive and memory B cells from the same patients. The analysis of an independent single-cell B-cell receptor sequencing (scBCR-seq) dataset of DENV-infected patients revealed that the peripheral blood PCs and PBs exhibited the highest clonal expansion in secondary DENV infection compared to other B cell subsets. These clonally expanded cells also expressed the highest levels of tissue-homing genes, including SELPLG. In addition, by utilizing a public scRNA-seq dataset of SARS-CoV2 infection, we demonstrated the upregulation of several tissue-homing genes in PCs and PBs. Our study provides evidence for the potential roles of tissue-homing B cell subsets in the context of immune responses against viral infections in humans.
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    Metal Oxide Nanostructures Enhanced Microfluidic Platform for Efficient and Sensitive Immunofluorescence Detection of Dengue Virus
    (2023-11-01)
    Pormrungruang, Pareesa
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    Phanthanawiboon, Supranee
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    Jessadaluk, Sukittaya
    ;
    Larpthavee, Preeda
    ;
    Thaosing, Jiraphon
    Rapid and sensitive detection of Dengue virus remains a critical challenge in global public health. This study presents the development and evaluation of a Zinc Oxide nanorod (ZnO NR)-surface-integrated microfluidic platform for the early detection of Dengue virus. Utilizing a seed-assisted hydrothermal synthesis method, high-purity ZnO NRs were synthesized, characterized by their hexagonal wurtzite structure and a high surface-to-volume ratio, offering abundant binding sites for bioconjugation. Further, a comparative analysis demonstrated that the ZnO NR substrate outperformed traditional bare glass substrates in functionalization efficiency with 4G2 monoclonal antibody (mAb). Subsequent optimization of the functionalization process identified 4% (3-Glycidyloxypropyl)trimethoxysilane (GPTMS) as the most effective surface modifier. The integration of this substrate within a herringbone-structured microfluidic platform resulted in a robust device for immunofluorescence detection of DENV-3. The limit of detection (LOD) for DENV-3 was observed to be as low as 3.1 × 10<sup>−4</sup> ng/mL, highlighting the remarkable sensitivity of the ZnO NR-integrated microfluidic device. This study emphasizes the potential of ZnO NRs and the developed microfluidic platform for the early detection of DENV-3, with possible expansion to other biological targets, hence paving the way for enhanced public health responses and improved disease management strategies.
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    Individual, Household, and Community Drivers of Dengue Virus Infection Risk in Kamphaeng Phet Province, Thailand
    (2022-10-15)
    Ribeiro Dos Santos, Gabriel
    ;
    Buddhari, Darunee
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    Iamsirithaworn, Sopon
    ;
    Khampaen, Direk
    ;
    Ponlawat, Alongkot
    Background: Dengue virus (DENV) often circulates endemically. In such settings with high levels of transmission, it remains unclear whether there are risk factors that alter individual infection risk. Methods: We tested blood taken from individuals living in multigenerational households in Kamphaeng Phet province, Thailand for DENV antibodies (N = 2364, mean age 31 years). Seropositivity ranged from 45.4% among those 1-5 years old to 99.5% for those >30 years. Using spatially explicit catalytic models, we estimated that 11.8% of the susceptible population gets infected annually. Results: We found that 37.5% of the variance in seropositivity was explained by unmeasured household-level effects with only 4.2% explained by spatial differences between households. The serostatus of individuals from the same household remained significantly correlated even when separated by up to 15 years in age. Conclusions: These findings show that despite highly endemic transmission, persistent differences in infection risk exist across households, the reasons for which remain unclear.
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    Plasmid DNA encoding neutralizing human monoclonal antibody without enhancing activity protects against dengue virus infection in mice
    (2021-07-01)
    Benjathummarak, Surachet
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    Yamanaka, Atsushi
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    Krasae, Thanyaluk
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    Pipattanaboon, Chonlatip
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    Injampa, Subenya
    Objective: To evaluate the expression of DNA plasmid-harboring modified antibody gene that produces neutralizing human monoclonal antibodies against four serotypes of dengue virus (DENV) without enhancing activity in BALB/c mice. Methods: We constructed pFUSE-based vectors (pFUSE-1G7C2-hVH and pFUSE-1G7C2-hVL) containing genes encoding the variable domains of the heavy or light chain of the anti-dengue virus antibody 1G7C2, a human IgG1 that has been characterized for its neutralizing activity to DENV-1-4. Leucine (L) at positions 234 and 235 on the Fc CH2 domain in pFUSE-1G7C2-hVH was mutated to alanine (A) (LALA mutation) by site direct mutagenesis, and the new plasmid was termed pFUSE-1G7C2-hVH-LALA. An equal amount of pFUSE-1G7C2-hVL and 1G7C2hG1-LALA plasmids were co-Transfected into Chinese hamster ovary cells (CHO-K1) and a single dose of 100 μg 1G7C2-hG1-LALA plasmid was intramuscularly injected, followed by electroporation in BALB/c mice. The secreted 1G7C2-hG1-LALA antibodies in cell culture supernatant and mouse serum were examined for their biological functions, neutralization and enhancing activity. Results: The co-Transfection of heavy-And light-chain 1G7C2-hG1-LALA plasmids in CHO-K1 cells produced approximately 3 900 ng/mL human IgG and neutralized 90%-100% all four DENV, with no enhancing activity. Furthermore, the modified human IgG was produced more than 1 000 ng/mL in mouse serum on day 7 post plasmid injection and showed cross-neutralization to four DENV serotypes. Subsequently, antibody production and neutralization decreased rapidly. Nevertheless, the secreted neutralizing 1G7C2-hG1-LALA in mouse serum demonstrated complete absence of enhancing activities to all DENV serotypes. Conclusions: These findings reveal that a new modified 1G7C2-hG1-LALA expressing plasmid based on gene transfer is a possible therapeutic antibody candidate against DENV infection.