title: Establishment and Comparison of Two Different Diagnostic Platforms for Detection of DENV1 NS1 Protein.
abstract: Dengue virus (DENV) infection is currently at pandemic levels, with populations in tropical and subtropical regions at greatest risk of infection. Early diagnosis and management remain the cornerstone for good clinical outcomes, thus efficient and accurate diagnostic technology in the early stage of the disease is urgently needed. Serotype-specific monoclonal antibodies (mAbs) against the DENV1 nonstructural protein 1 (NS1), DA12-4, DA13-2, and DA15-3, which were recently generated using the hybridoma technique, are suitable for use in diagnostic platforms. Immunofluorescence assay (IFA), enzyme-linked immunosorbent assay (ELISA) and Western blot analysis further confirmed the serotype specificity of these three monoclonal antibodies. The ELISA-based diagnostic platform was established using the combination of two highly sensitive mAbs (DA15-3 and DB20-6). The same combination was also used for the flow cytometry-based diagnostic platform. We report here the detection limits of flow cytometry-based and ELISA-based diagnostic platforms using these mAbs to be 0.1 and 1 ng/mL, respectively. The collected clinical patient serum samples were also assayed by these two serotyping diagnostic platforms. The sensitivity and specificity for detecting NS1 protein of DENV1 are 90% and 96%, respectively. The accuracy of our platform for testing clinical samples is more advanced than that of the two commercial NS1 diagnostic platforms. In conclusion, our platforms are suitable for the early detection of NS1 protein in DENV1 infected patients.
authors.affiliation: Graduate Institute of Life Sciences, National Defense Medical Center, Taipei 114, Taiwan. bigerliang@yahoo.com.tw.|Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan. bigerliang@yahoo.com.tw.
authors.forename: Yin-Liang
authors.initials: YL
authors.lastname: Tang
authors.affiliation: Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan. chienyu412@gmail.com.
authors.forename: Chien-Yu
authors.initials: CY
authors.lastname: Chiu
authors.affiliation: Division of Infectious Diseases, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan. infectionman@gmail.com.|School of Medicine, Graduate Institute of Medicine, Sepsis Research Center, Kaohsiung Medical University, Kaohsiung 807, Taiwan. infectionman@gmail.com.
authors.forename: Chun-Yu
authors.initials: CY
authors.lastname: Lin
authors.affiliation: Division of Infectious Diseases, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan. locusthao@gmail.com.|School of Medicine, Graduate Institute of Medicine, Sepsis Research Center, Kaohsiung Medical University, Kaohsiung 807, Taiwan. locusthao@gmail.com.
authors.forename: Chung-Hao
authors.initials: CH
authors.lastname: Huang
authors.affiliation: Division of Infectious Diseases, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan. infchen@gmail.com.|School of Medicine, Graduate Institute of Medicine, Sepsis Research Center, Kaohsiung Medical University, Kaohsiung 807, Taiwan. infchen@gmail.com.|Department of Biological Science and Technology, College of Biological Science and Technology, National Chiao Tung University, Hsinchu 300, Taiwan. infchen@gmail.com.
authors.forename: Yen-Hsu
authors.initials: YH
authors.lastname: Chen
authors.affiliation: Institute of Molecular Biology and Biotechnology, National Institutes of Health and Philippine Genome Center, University of the Philippines, Manila 1000, Philippines. raul.destura@biotechmanila.upm.edu.ph.
authors.forename: Raul V
authors.initials: RV
authors.lastname: Destura
authors.affiliation: Graduate Institute of Microbiology and Public Health, College of Veterinary Medicine, National Chung-Hsing University, Taichung 402, Taiwan. dychao@dragon.nchu.edu.tw.
authors.forename: Day-Yu
authors.initials: DY
authors.lastname: Chao
authors.affiliation: Graduate Institute of Life Sciences, National Defense Medical Center, Taipei 114, Taiwan. hcw0928@gate.sinica.edu.tw.|Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan. hcw0928@gate.sinica.edu.tw.
authors.forename: Han-Chung
authors.initials: HC
authors.lastname: Wu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D003715:Dengue / Q000175:diagnosis* / Q000276:immunology*; D003716:Dengue Virus / Q000145:classification / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D005260:Female; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards; D006801:Humans; D051379:Mice; D011994:Recombinant Proteins; D012680:Sensitivity and Specificity; D012703:Serotyping; D017361:Viral Nonstructural Proteins / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; C082515:NS1 protein, dengue-1 virus; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins
keywords: dengue virus; diagnosis; monoclonal antibody; nonstructural protein 1