標題: Establishment and Comparison of Two Different Diagnostic Platforms for Detection of DENV1 NS1 Protein
作者: Tang, Yin-Liang
Chiu, Chien-Yu
Lin, Chun-Yu
Huang, Chung-Hao
Chen, Yen-Hsu
Destura, Raul V.
Chao, Day-Yu
Wu, Han-Chung
生物科技學系
Department of Biological Science and Technology
關鍵字: dengue virus;nonstructural protein 1;monoclonal antibody;diagnosis
公開日期: 1-Nov-2015
摘要: 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.
URI: http://dx.doi.org/10.3390/ijms161126069
http://hdl.handle.net/11536/129392
ISSN: 1422-0067
DOI: 10.3390/ijms161126069
期刊: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume: 16
Issue: 11
起始頁: 27850
結束頁: 27864
Appears in Collections:Articles