Abstract
Dengue virus (DENV) nonstructural 1 (NS1) protein is a specific and sensitive biomarker for the diagnosis of dengue. In this study, an efficient electrochemical biosensor that uses chemically modified affinity peptides was developed for the detection of dengue virus NS1. A series of amino acid-substituted synthetic peptides was rationally designed, chemically synthesized and covalently immobilized to a gold sensor surface. The sensor performance was monitored via square wave voltammetry (SWV) and electrochemical impedance spectroscopy (EIS). Potential affinity peptides specific for NS1 were chosen according to the dynamic current decrease in SWV experiments. Using circular dichroism, the molar ellipticity of peptides (DGV BP1–BP5) was determined, indicating that they had a mostly similar in random coil structure, not totally identical. Using SWV, DGV BP1 was selected as a promising recognition peptide and limit of detection for NS1 was found to be 1.49 μg/mL by the 3-sigma rule. DGV BP1 showed good specificity and stability for NS1, with low signal interference. The validation of the sensor to detect NS1 proteins was confirmed with four dengue virus culture broth (from serotype 1 to 4) as proof-of-concept. The detection performance of our sensor incorporating DGV BP1 peptides showed a statistically significant difference. These results indicate that this strategy can potentially be used to detect the dengue virus antigen, NS1, and to diagnosis dengue fever within a miniaturized portable device in point-of-care testing.
Highlights
Dengue fever is a one of the major disease that inflicts a heavy global public health burden [1,2,3]
Rational design and synthesis of affinity peptides Our preliminary results indicated that phage-displayed peptides were specific for nonstructural 1 (NS1) [5], we assumed that free peptides, separated from the M13 phages, may be suitable to use for the creation of an electrochemical sensor for the efficient detection of Dengue virus (DENV) antigens, NS1
One limitation of the previous study is that the interaction of the peptides-displayed on M13 phage particles with NS1 targets may be different when affinity peptides are free in solution or when affinity peptides are immobilized on a surface with varying densities
Summary
Dengue fever is a one of the major disease that inflicts a heavy global public health burden [1,2,3]. The appearance of dengue fever derives from mosquito-borne virus, and it can spread globally into many countries with a tremendous increase in epidemic and endemic proportions [4]. Clinical symptoms and illness showing dengue-induced shock syndrome appear rapidly after viral infection [2, 3, 5].
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