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    Item type:Publication,
    Laser-induced graphene electrochemical immunosensors for rapid and sensitive serological detection: A case study on dengue detection platform
    (2025-06-01)
    Inlumphan, Supawee
    ;
    Wongwiriyapan, Winadda
    ;
    Khemasiri, Narathon
    ;
    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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    Item type:Publication,
    Urinary dengue NS1 detection on Au-decorated ZnO nanowire platform
    (2024-06-15)
    Sitthisuwannakul, Kannika
    ;
    Sukthai, Ratchanon
    ;
    Zhu, Zetao
    ;
    Nagashima, Kazuki
    ;
    Chattrairat, Kunanon
    Biodetection for non-invasive diagnostics of fluids, especially urine, remains a challenge to scientists due to low target concentrations. And biological complexes of the detection target may contain contaminants that also interfere with any assay. Dengue non-structural 1 protein (Dengue NS1) is an important biomarker for dengue hemorrhagic fever and dengue shock syndrome. Here, we developed an Au-decorated nanowire platform and applied it with a sandwich fluorophore-linked immunosorbent well plate assay (FLISA) to detect Dengue NS1 in urine. For the platform, we fabricated zinc oxide (ZnO) nanowires to provide a high surface area and then coated them with gold nanoparticles (ZnO/Au nanowires) to simply modify the Dengue NS1 antibody and enhance the fluorescence intensity. Our platform employs a sandwich FLISA that exhibits high sensitivity, specifically detecting Dengue NS1 with a limit of detection (LOD) of 1.35 pg/mL. This LOD was 4500-fold lower than the LOD of a commercially available kit for Dengue NS1 enzyme-linked immunosorbent assay. We believe that our ZnO/Au nanowire platform has the potential to revolutionize the field of non-invasive diagnostics for dengue.
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    Item type:Publication,
    Metal Oxide Nanostructures Enhanced Microfluidic Platform for Efficient and Sensitive Immunofluorescence Detection of Dengue Virus
    (2023-11-01)
    Pormrungruang, Pareesa
    ;
    Phanthanawiboon, Supranee
    ;
    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.