Abstract
Rational selection of predicted peptides to be employed as templates in molecular imprinting was carried out for the heat-denatured non-structural protein 1 (NS1) of dengue virus (DENV). Conservation analysis among 301 sequences of Brazilian isolates of DENV and zika virus (ZIKV) NS1 was carried out by UniProtKB, and peptide selection was based on in silico data of the conservational, structural and immunogenic properties of the sequences. The selected peptide (from dengue 1 NS1) was synthesized and employed as a template in the electropolymerization of polyaminophenol-imprinted films on the surface of carbon screen-printed electrodes. Heat denaturation of the protein was carried out prior to analysis, in order to expose its internal hidden epitopes. After removal of the template, the molecularly imprinted cavities were able to rebind to the whole denatured protein as determined by electrochemical impedance spectroscopy. This label-free sensor was efficient to distinguish the NS1 of DENV from the NS1 of ZIKV. Additionally, the sensor was also selective for dengue NS1, in comparison with human serum immunoglobulin G and human serum albumin. Additionally, the device was able to detect the DENV NS1 at concentrations from 50 to 200 μg L−1 (RSD below 5.04%, r = 0.9678) in diluted human serum samples. The calculated LOD and LOQ were, respectively, 29.3 and 88.7 μg L−1 and each sensor could be used for six sequential cycles with the same performance.
Highlights
The genus Flavivirus contains more than 70 different viruses classi fied in 53 species
Epitope-imprinted films were deposited on carbon screen-printed electrodes (C-SPEs) by electro polymerization using a mixture of 2.5 mmol L− 1 3-AP and 250 μmol L− 1 28WTEQYKFQA36 peptide, prepared in 10 mmol L− 1 phosphate-buffered saline (PBS) at pH 7.4
The combination of these techniques provides a versatile protocol which can be applied for epitope imprinting for various molecularly imprinted polymers (MIPs) applications
Summary
The genus Flavivirus contains more than 70 different viruses classi fied in 53 species. Japanese encephalitis virus, dengue virus (DENV) and zika virus (ZIKV) are the most frequent human infections caused by these vector-borne diseases. Dengue is a vector-borne disease transmitted to a human host through the bite of an Aedes aegypti or. Langerak et al (2019) discussed the occurrence of an antibody-dependent enhancement between DENV and ZIKV due to the cross-reactivity of the host antibodies. This cross-reactivity occurs due to the high degree of homology observed between these species. The first licensed dengue vaccine, CYD-TDV, still offers risks for seronegative patients and this lack of preventive measures emphasizes the need for a precise diagnosis (Wilder-Smith, 2018)
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