DNA aptamer selection and construction of an aptasensor based on graphene FETs for Zika virus NS1 protein detection

Abstract

Zika virus (ZIKV) is a mosquito-borne virus that is phylogenetically close to other medically important flaviviruses with high global public health significance, such as dengue (DENV) and yellow fever (YFV) viruses. Correct diagnosis of a flavivirus infection can be challenging, particularly in world regions where more than one flavivirus co-circulates and YFV vaccination is mandatory. Acid nucleic aptamers are oligonucleotides that bind to a specific target molecule with high affinity and specificity. Because of their unique characteristics, aptamers are promising tools for biosensor development. Aptamers are usually obtained through a procedure called “systematic evolution of ligands by exponential enrichment” (SELEX). In this study, we select an aptamer (termed ZIKV60) by capillary electrophoresis SELEX (CE-SELEX) to the Zika virus non-structural protein 1 (NS1) and counterselection against the NS1 proteins of DENV (serotypes 1, 2, 3, and 4) and YFV. The ZIKV60 dissociation constant (Kd) is determined by enzyme-linked oligonucleotide assay (ELONA) and the aptamer specificity is evaluated by quantitative real-time polymerase chain reaction. ZIKV60 shows a high binding affinity to the ZIKV NS1 protein with a Kd value of 2.28 ± 0.28 nM. The aptamer presents high specificity for ZIKV NS1 compared to NS1 of DENV and YFV. Furthermore, graphene field-effect transistor devices functionalized with ZIKV60 exhibit an evident identification of NS1 protein diluted in human serum. These results point to the applicability of biosensors based on the ZIKV60 aptamer for the differential diagnosis of the Zika virus.

Aptamer–Protein Interactions: From Regulation to Biomolecular Detection

“…Subsequently, ELAA has been extensively studied to detect aptamer–protein interactions. Almeida et al selected aptamers against zika virus nonstructural protein 1 (ZIKVNS1), termed ZIKV60, and performed ELAA to analyze ZikV60-ZikVNS1 interactions . Specifically, ZIKVNS1 was incubated with horseradish peroxidase (HRP)-labeled ZIKV60 at different concentrations, followed by adding 3,5,3′,5′-tetramethylbenzidine (TMB) to initiate colorimetric reactions.…”

“…Almeida et al selected aptamers against zika virus nonstructural protein 1 (ZIKVNS1), termed ZIKV60, and performed ELAA to analyze ZikV60-ZikVNS1 interactions. 272 Specifically, ZIKVNS1 was incubated with horseradish peroxidase (HRP)-labeled ZIKV60 at different concentrations, followed by adding 3,5,3′,5′-tetramethylbenzidine (TMB) to initiate colorimetric reactions. Based on measurements of absorbance at 450 nm, the saturation−binding curve was plotted and showed that ZIKV60 presented a high binding affinity to ZIKVNS1 with a K D value of 2.28 ± 0.28 nM.…”

Ultrafast and highly sensitive detection of SARS-CoV-2 spike protein by field-effect transistor graphene-based biosensors

“…However, while these rapid tests provide reliable diagnosis after the first days of symptoms, they are inappropriate for clinical decision-making in the early stages of the disease [12]. Conversely, the association of graphene with advanced functional materials, such as monoclonal antibodies and aptamers, for instance, represents a promising route towards the development of next-generation medical platforms for highly sensitive antigen detection [13][14][15][16]. Given its two-dimensional nature and high carrier mobility, graphene is highly responsive to interactions with external agents [17].…”

Recent Developments in DNA-Nanotechnology-Powered Biosensors for Zika/Dengue Virus Molecular Diagnostics

“…In addition, two-dimensional and three-dimensional DNA nanostructures can encapsulate vaccine antigens and biochemical molecules [ 39 ]. Moreover, they have a number of advantages, such as the reduction of drug toxicity and enhancement of drug targeting, and have high potential as powerful and simple design techniques for the self-assembly of nanostructures [ 45 ]. Among these, the most noteworthy is the DNA-nanotechnology-based biosensor.…”