2. Academic Validation
  3. Development of a sensitive sandwich ELISA for the envelope domain III protein of dengue virus type 2

Development of a sensitive sandwich ELISA for the envelope domain III protein of dengue virus type 2

  • J Immunol Methods. 2025 Dec:545:114000. doi: 10.1016/j.jim.2025.114000.
Ziting Ma 1,  Jingyi Su 1,  Zelin Han 2,  Ximing Ren 1,  Jinnian Guo 1,  Lu Jiang 3,  Suqing Zhao 4
Affiliations

Affiliations

  • 1 Department of Pharmaceutical Engineering, School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou, Guangdong 510006, People's Republic of China.
  • 2 Department of Biomedical Engineering, School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou, Guangdong 510006, People's Republic of China.
  • 3 Department of Biomedical Engineering, School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou, Guangdong 510006, People's Republic of China. Electronic address: [email protected].
  • 4 Department of Pharmaceutical Engineering, School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou, Guangdong 510006, People's Republic of China. Electronic address: [email protected].
PMID: 41183705 DOI: 10.1016/j.jim.2025.114000
Abstract

Dengue, caused by the Dengue Virus (DENV), is a rapidly spreading infectious disease that poses a significant global health challenge. Rapid and accurate diagnosis is critical for effective disease management and epidemic control, as dengue is a kind of vector-borne disease and lacks definitive medical treatments. Current antibodies and antigens for DENV Immunoassay, including immunoglobulin G (IgG), immunoglobulin M (IgM), and non-structural protein 1 (NS1), have drawbacks like cross-reaction and false-negative results in the late stages of primary Infection or secondary Infection. Envelope domain III (EDIII) protein, which is immunogenic and is a recognition region and binding site for DENV receptors, can also be the potential detection target of DENV Infection. However, few studies of EDIII protein as a detection target have been reported. Therefore, this study developed a sensitive and specific ELISA for DENV-2_EDIII protein detection. Specific polyclonal antibodies (pAbs) against the DENV-2_EDIII protein were generated by immunizing the New Zealand White rabbits with the recombinant DENV-2_EDIII protein. Using these pAbs, a highly sensitive and specific ELISA method was developed. The assay demonstrated a wide detection range (7.8 ng/mL to 500 ng/mL) and a low limit of detection (LOD) of 0.156 ng/mL and showed no cross-reactivity with Other orthoflaviviruses. These findings may provide a new methodological framework and research direction for the diagnosis of dengue after further clinical verifications.

Keywords

Dengue virus type 2 (DENV-2); Envelope domain III (EDIII); Enzyme-linked immunosorbent assay (ELISA); Immunoassay; Polyclonal antibody (pAb).

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