期刊
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
卷 149, 期 -, 页码 591-602出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jpba.2017.11.064
关键词
Optical; Immunosensor; Immunofluorescence; Dengue virus; NS1
资金
- University of Malaya Research Grant [UMRG] [RP012C-14SUS]
- Fundamental Research Grant Scheme [FRGS] from the Ministry of Higher Education of Malaysia [MOHE] [FP041-2016]
- University of Malaya Postgraduate Research Grant [PG120-2012B]
The sharp increase in incidence of dengue infection has necessitated the development of methods for the rapid diagnosis of this deadly disease. Here we report the design and development of a reliable, sensitive, and specific optical immunosensor for the detection of the dengue nonstructural protein 1 (NS1) biomarker in clinical samples obtained during early stages of infection. The present optical NS1 immunosensor comprises a biosensing surface consisting of specific monoclonal NS1 antibody for immunofluorescence-based NS1 antigen determination using fluorescein isothiocyanate (FITC) conjugated to IgG antibody. The linear range of the optical immunosensor was from 15 - 500 ngmL(-1), with coefficient of determination (R-2) of 0.92, high reproducibility (the relative standard deviation obtained was 2%), good stability for 21 days at 4 degrees, and low detection limit (LOD) at 15 ng mL(-1). Furthermore, the optical immunosensor was capable of detecting NS1 analytes in plasma specimens from patients infected with the dengue virus, with low cross - reaction with plasma specimens containing the Japanese encephalitis virus (AV) and Zika virus. No studies have been performed on the reproducibility and cross -reactivity regarding NS1 specificity, which is thus a limitation for optical NS1 immunosensors. In contrast, the present study addressed these limitations carefully where these two important experiments were conducted to showcase the robustness of our newly developed optical-based fluorescence immunosensor, which can be practically used for direct NS1 determination in any untreated clinical sample. (C) 2017 Elsevier B.V. All rights reserved.
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