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Dutta, R.; Rajendran, K.; Jana, S.K.; Saleena, L.M.; Ghorai, S. Extensive Use of Graphene in Detection of Viruses. Encyclopedia. Available online: (accessed on 19 April 2024).
Dutta R, Rajendran K, Jana SK, Saleena LM, Ghorai S. Extensive Use of Graphene in Detection of Viruses. Encyclopedia. Available at: Accessed April 19, 2024.
Dutta, Reshmi, Kokilavani Rajendran, Saikat Kumar Jana, Lilly M. Saleena, Suvankar Ghorai. "Extensive Use of Graphene in Detection of Viruses" Encyclopedia, (accessed April 19, 2024).
Dutta, R., Rajendran, K., Jana, S.K., Saleena, L.M., & Ghorai, S. (2023, March 29). Extensive Use of Graphene in Detection of Viruses. In Encyclopedia.
Dutta, Reshmi, et al. "Extensive Use of Graphene in Detection of Viruses." Encyclopedia. Web. 29 March, 2023.
Extensive Use of Graphene in Detection of Viruses

Graphene and its derivatives offer considerable merits in biosensing applications for pathogenic virus detection.

graphene diagnostics graphene oxide

1. Introduction

Dengue virus (DENV), a member of the family Flaviviridae, is a single-stranded positive-sense RNA virus with a genome of 11 kb [1] and is transmitted by the female Aedes aegyptii mosquito vector [2]. DENV consists of three structural proteins, namely the membrane-associated protein M, core protein C, and envelope protein E, which are responsible for shaping the structure of the virus. In addition, it contains seven non-structural proteins, NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5, which play a role in the replication of the virus and many more cellular functions. The different serotypes of dengue are DENV-1, DENV-2, DENV-3, and DENV-4. Infection caused due to any of these serotypes is usually asymptomatic or flu-like symptoms develop, which might lead to severe dengue hemorrhagic fever or dengue shock syndrome [1]. A global study showed that about 390 million people get infected by dengue annually, of which 96 million exhibit clinical manifestations [2]. As of 23 September 2022, 110,473 dengue cases have been reported in India [3].
Conventional methods for dengue detection are laborious, time-consuming, costly and require skilled professionals. Hence, nanomaterial-based sensors can be a good option for dengue detection as they surpass all the issues mentioned above that conventional methods face. Nanomaterials have exceptional qualities, making them an ideal platform for usage in biosensors. They can be used for certain purposes by regulating physical and chemical properties, namely size, morphology, surface charge and solubility [4]. Such properties make nanomaterials ideal for their utility in biosensors to enhance target-specific reactions which respond to biochemical reactions such as temperature, pH, and the existence of enzymes [5]. Nanomaterials might boost the sensitivity of present diagnostic techniques due to features like high reactivity, adsorption capacity, particle size, and physicochemical bonding capacity. Utilizing nanoparticles might lead to enhancements in technology regarding time, specificity, mobility and convenience [4].
Graphene is one such material which is being used extensively in the development of sensors. Graphene is a premium material formed by a single-thick layer of sp2 hybridized carbon atoms arranged in a honeycomb lattice [6]. It is an extremely thin material and occurs as a few layers of graphite [7]. It displays outstanding electrochemical properties such as high thermal conductivity (above 3000 W m K−1) and low redox potential (−0.5 V–1.2) [8]. Optical absorption in the infrared region, a high surface area (2630 m2/g), completely impermeable to gas [9], and mechanical strength (about 1100 GPa) [10], are some of the properties for which graphene is used for diagnostic purposes [9][10]. The different forms of graphene include graphene oxide (GO), reduced graphene oxide (rGO), graphene sheets [7], and layered graphenes such as a few layered graphenes and multilayered graphene [7][11]. Some of the widely studied derivatives of graphene are GO and rGO (Figure 1). They display properties similar to graphene, such as flexibility, transparency, and low cytotoxicity. The hydrophilic nature and presence of reactive functional groups make them an ideal material to be used in a biosensor [12]. The hydrophilic feature plays a key role in assembling the biosensor as it permits film preparation by drop casting, spin coating, ink-jet printing, and electrode material processing. Research on these materials has successfully led to their use in different fields such as electronics, optics, sensors, and filtration [8]. The synthesis of GO involves vigorous oxidation of graphite–graphene solutions and hence leads to the production of a complex material having a variety of functional groups whose properties and quantity rely on the method of preparation. There is a tendency for functional groups of GO to get modified on the surface due to the attachment of redox species such as enzymes, peptides, or DNA and RNA [13]. In an acidic medium, graphite is oxidized to GO using Hummers’ and Offemans’ methods [14]. GO’s disrupted sp2 hybridized bonding is responsible for its electrically insulating nature, however, GO can also be a good semiconductor when oxidized extensively [15].
Figure 1. Structures of graphene (G), graphene oxide (GO), and reduced graphene oxide (rGO) (Reused with permission from Nanomaterials, MDPI [16]). Aromaticity in graphene is local, with two π-electrons located over every hexagon ring [17].
When graphene is converted to GO, it increases the hydrophilicity of the surface and results in the formation of big functional groups [18]. The presence of these oxygenated groups (hydroxyl and epoxy) thus aids in the formation of a stable dispersion in aqueous media and other polar solvents [14] and further enables biochemical and bioconjugation reactions to occur with ease [14][19]. GO is a remarkable adsorbant of proteins and antibodies, making it a useful biomaterial [20]. GO, on reduction, produces rGO by removing the oxidized functional groups of GO [21]. The reduction procedure establishes considerable changes involving surface properties in rGO [20]. The absence of most of the oxygen-containing functional groups from GO partly restores the sp2 structure [22] and improves the rGO conductivity [22][23]. Some unique properties of rGO involve conductivity, fluorescence quenching, peroxide-like enzyme activity, intrinsic Raman activity, oxidation-reduction capacity, and the ability to anchor different nanoparticles, enzymes, proteins, and nucleic acids [13]. While GO has inferior electrical properties, it is a better conductor and can retain its ability to disperse in water [24]. GO and rGO can be utilized in the preparation of a wide range of graphene-based nanocomposites. Some materials incorporated with graphene derivatives are nanoparticles, quantum dots, nanoclusters, polymers, and a variety of biomolecules [25]. Graphene and its derivatives undergo non-covalent interactions such as London forces, polarization, hydrophobic effects, and electrostatic force of attraction with adsorbates [26]. Such interactions confirm the adsorbate’s contact with a surface, leading to alterations in electronic characteristics that could be used in sensing. GO possesses a high loading ratio for biomolecules which can go up to 200% compared to other nanocarriers. This could be a useful feature for the development of sensitive electrochemical immunosensors [27]. A noteworthy point could be that 2D functionalized graphene derivatives and composites might undergo restacking. Restacking happens when multilayer Van der Waals materials are formed due to the powerful interaction of non-covalent interlayers. Developing a firm 3D network is crucial for yielding new and stable graphene nanocomposites that do not undergo restacking and possess beneficial features such as high surface area, tunable electronic features, improved conductivity and electrocatalytic characteristics, available inner space, and improved mechanical stability. Such features are suitable for systematic and selective biosensing [28].
Graphene and its derivatives offer considerable merits in biosensing applications for pathogenic virus detection. These materials are more conductive electrically and thermally, flexible, not heavy, biocompatible, and have a large surface area which is a gold standard for electrochemical platforms. Graphene, GO, and rGO successfully quench photoluminescence. Incorporating befitting nanomaterials into graphene and its derivatives can improve the quality of the known detection methods, e.g., Au–Ag nanoparticles transmit a notable surface-enhanced Raman scattering (SERS) signal. The sensor’s sensitivity is related to the transforming electronic features of graphene when the virus adsorbates are present. The large surface area of graphene-based materials helps the adsorption of analytes from the environment. When antibodies, aptamers, or nucleic acids are incorporated into graphene and its derivatives, they can bind to counterparts specific to a particular virus. This enhances the selectivity of a biosensor [13]. Therefore, biosensors utilizing graphene and its derivatives can be used efficiently for the detection of not only the dengue virus but also other upcoming pathogenic viruses.

2. Extensive Use of Graphene in the Detection of Viruses

The light, chemically stable, and conductive nature of graphene makes it a success in its use in the detection of viruses. The incorporation of functional groups into the hybrid structure leads to swift virus detection [13]. Graphene and its derivatives are often used in the detection of viruses due to their high specificity and less time expenditure. Polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) are thought to be sensitive techniques in the detection of the foot and mouth disease virus (FMDV) [29]. PCR has certain limitations, which lead to the generation of false positive results. A nano-PCR system, which uses graphene oxide–gold nanoparticles nanocomposites (GO-AuNPs), exhibits improved PCR efficiency [30].
In recent years rGO has found wide application in electrochemical biosensing for the detection of the H1N1 (haemagglutinin type 1 and neuraminidase Type 1) strain of the influenza virus. Label-free detection of the H1N1 virus was done using an electrochemical immunosensor coated with rGO and incorporated with a microfluidic platform [31]. The rGO obtained from shellac, used in label-free electrochemical biosensors, proved to be a cost-effective method for the detection of the influenza virus H1N1 [32].
Low-cost ultrasensitive biosensors using graphene and its derivatives exhibit excellent and promising results in the detection of human papilloma virus (HPV16) DNA in patients suffering from HPV16-positive head and neck cancer [33]. Reduced graphene oxide field effect transistors (rGO-FETs) are used for highly sensitive and selective detection of HPV16 E7 protein. To acquire selective sensing, specific probes are used on the conducting rGO channel of the transistor [34].
Sulfonated magnetic nanoparticles functionalized with reduced graphene oxide (SMRGO), having antiviral properties, were used to eliminate herpes simplex virus type 1 (HSV-1). Graphene has remarkable photothermal properties that inactivate the viruses captured with near-infrared (NIR) irradiation [35]. Graphene-mediated surface-enhanced Raman scattering (G-SERS) is often used to identify norovirus (NoV) in a sample of human faeces. In this unique biosensing system, magnetic derivative molybdenum trioxide nanocubes functioned as the SERS nanotag, and single-layer graphene oxide (SLGO) served as the signal reporter. With this technology, rapid signal amplification for the detection of NoV could be achieved [36].
Detection of the hepatitis C virus (HCV) could be made using a highly sensitive assay that involves the use of reduced graphene oxide nanosheets (rGONS) and a hybridization chain reaction (HCR) amplification technique. This method uses the selective adsorption capacity of rGONS to fluorophore probes with different conformations [37]. The enhanced fluorescence quenching capacity of rGO leads to inverse upregulation of the assay sensitivity by a reduction in the background signal of the probes [38][39][40]. The methods of detecting different viruses using graphene are listed in Table 1.
Table 1. Graphene-based detection for different viruses. (H1N1—haemagglutinin type 1 and neuraminidase type 1 influenza virus, HPV—human papilloma virus, HSV1—herpes simplex virus 1, NoV—norovirus, HCV—hepatitis C virus).


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