Electrochemical Immunosensor Based on Antibody-Nanoparticle Hybrid for Specific Detection of the Dengue Virus NS1 Biomarker

Abstract

Dengue virus infection is a very deadly disease that threatens human lives in subtropical and tropical regions. Thus, for early and reliable diagnosis of Dengue virus infection, a sensitive, specific, and label-free electrochemical immunosensor was developed for the direct detection of the unstructured protein NS1. The NS1 biosensor was designed with a biosensing surface consisting of antifouling moieties and biorecognition molecules to enhance the specificity of the immunosensor for target analyte detection in complicated biological samples such as human sera. The immunosensor exhibited a wide detection range (5-4000 ng mL −1 ) based on electrochemical impedance spectroscopy (EIS) measurements with a coefficient of determination (R 2 ) of 0.94, correlation coefficient (R) of 0.95, high reproducibility, and good stability for 21 days at 4 • C. The developed immunosensor was able to detect the NS1 antigen in actual serum specimens from patients infected with Dengue virus. Moreover, the immunosensor is not only highly selective for the NS1 antigen, but also did not cross-react with human sera infected with malaria parasites.

The capacitive sensing of NS1 Flavivirus biomarker

“…5a) with a sensitivity of~22% decade −1 , a RSD (%) of 7 ± 4 ( Fig. 5a) and a LoD of 0.5 ng mL −1 , thus approximately sixty-fold lower than those obtained by EIS-ES and ten-fold lower than an immunosensor based on EIS reported in the literature by others (Darwish et al, 2016). In addition, the repeatability of the NS1 biosensors was assessed using five interday and five intra-day replicate measurements of a 50 ng mL −1 standard solution in neat serum wherein the coefficients of variation obtained were 7% and 8%, for ECS, while values of 6% and 3% were achieved by EIS.…”

Molecular Biosensors for Electrochemical Detection of Infectious Pathogens in Liquid Biopsies: Current Trends and Challenges

“…Regarding viral infection determination [ 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 ], electrochemical biosensing approaches have been focused mainly on the detection of: the whole virus such as human enterovirus 71 [ 55 ]; specific viral proteins (p24 from HIV [ 46 , 47 , 48 , 49 ] and NS1 from Dengue [ 54 ]); autoantibodies against specific viral antigens (pseudorabies virus [ 50 ], HIV-1 and HIV-2 [ 51 , 56 ] and hemagglutinin, HA, from avian influenza virus H5N1 [ 52 , 53 ]) and characteristic DNA sequences (Dengue virus serotype 3, DENV-3, [ 57 ] and high-risk human papillomavirus, hrHPV, strains HPV-16 and HPV-18 [ 58 ]). …”

Electrochemical Biosensing for the Diagnosis of Viral Infections and Tropical Diseases

“…A sensitive, specific, and label‐free electrochemical immunosensor was developed for early and reliable diagnosis of dengue virus infection though the determination of the unstructured protein NS1 secreted during the early phase of the disease with a concentration of up to 50 μg mL −1 . The immunosensing scaffold employed ITO electrodes modified by in situ generated aryl diazonium cations derived zwitterionic antifouling molecules, AuNPs and monoclonal antibodies.…”