NS1 glycoprotein detection in serum and urine as an electrochemical screening immunosensor for dengue and Zika virus

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作者
Priscila D. Mendonça
Lorenna K. B. Santos
Marcos V. Foguel
Marco A. B. Rodrigues
Marli T. Cordeiro
Luís M. Gonçalves
Ernesto T. A. Marques
Rosa F. Dutra
机构
[1] Federal University of Pernambuco,Biomedical Engineering Laboratory
[2] Federal University of Pernambuco,Electronic Department
[3] Aggeu Magalhães Institute,Department of Virology and Experimental Therapy, Oswaldo Cruz Foundation – FIOCRUZ
[4] University of São Paulo,Institute of Chemistry
[5] University of Pittsburgh,Department of Infectious Diseases and Microbiology
[6] Center for Vaccine Research,undefined
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关键词
Immunosensor; Label-free; NS1; Dengue virus; Zika virus;
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摘要
The incidence of infection by the dengue virus (DENV) has grown dramatically, reaching 128 countries in tropical and subtropical regions worldwide, with a pattern of hyper-endemicity. DENV is a mosquito-borne disease having four serotypes, one or two circulating in epidemic outbreaks. The diagnosis of DENV is challenging mainly due to the circulation of new viruses with remarkable similarities, such as Zika (ZIKV) that may cause fetal microcephaly. DENV affects 390 million people per year, but these numbers may be higher due to the underreported and misclassified cases. Recently, the NS1 nonstructural protein has been described in serum and urine of DENV and ZIKV patients, suggesting its use as a biomarker for screening since a negative NS1 sample confirms the absence of these infections. Herein, a label-free immunosensor comprising an assembled nanostructured thin film of carbon nanotube-ethylenediamine is described. The advantage of in situ electrosynthesis of polymer film is to allow major control of thickness and conductivity, in addition to designing the reactive groups for functionalization. A quartz crystal microbalance system was used to estimate the thickness of the polymeric film obtained. The anti-NS1 monoclonal antibodies were immobilized to carbon nanotubes by covalent linkage, permitting a high stability during measurements. Analytical responses to NS1 were obtained by differential pulse voltammetry (DPV), showing a linear range from 20 to 800 ng mL−1 and reproducibility of 3.0%, with a limit of detection (LOD) of 6.8 ng mL- 1. This immunosensor was capable of detecting ZIKV and DENV NS1 in spiked urine and real serum in a clinical range.
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页码:4873 / 4885
页数:12
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