期刊
BIOSENSORS & BIOELECTRONICS
卷 191, 期 -, 页码 -出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2021.113419
关键词
DENV; NS1; Molecularly imprinted polymers; Sensor; Epitope; Label-free
类别
资金
- Fundacao de Amparo a Pesquisa do Estado de Minas Gerais (FAPEMIG, Belo Horizonte, Brazil) [CDS-APQ-00638-17]
- Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq, Brasilia, Brazil) [301564/2019-1, 427365/2018-0]
- Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES) [88881.133746/2016-01]
Based on conservation analysis, a peptide from dengue virus NS1 was selected as a template for molecular imprinting, resulting in a label-free sensor capable of distinguishing DENV from ZIKV. The sensor showed high selectivity and stability in detecting DENV NS1 in human serum samples.
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 mu g 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 mu g L-1 and each sensor could be used for six sequential cycles with the same performance.
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