key: cord-0299474-ii4nittr authors: Sengupta, Joydip; Adhikari, Arpita; Hussain, Chaudhery Mustansar title: Graphene-based analytical lab-on-chip devices for detection of viruses: A review date: 2021-06-24 journal: nan DOI: 10.1016/j.cartre.2021.100072 sha: 6d7d1dc321a5e06fe7e2c22bee562baa2e9d4eb9 doc_id: 299474 cord_uid: ii4nittr Lab on a chip (LOC) device intakes fluid and makes it flow through the microchannels, to achieve rapid, highly sensitive and low-cost analysis with significant yield. Graphene has vast potential to be used in LOC devices owing to its remarkable and unique properties. A trustworthy, swift, inexpensive and facile detection scheme is of urgency due to the current situation of COVID-19 to break the chain of transmission and lab-on-chip based biosensor has materialized itself as a realistic solution for this purpose. The addition of Graphene has augmented the sensing capability of the LOC devices to a superior level. Recently, Graphene-based lab-on-chip type biosensor is effectively used for faithful detection of SARS-CoV-2 and this draws the attention of the researchers to review the recent progress in Graphene-based LOC platforms for detection of viruses and the same has been reviewed here. to describe the miniaturized thin-film biosensors build-up by them. Afterwards, the term LOC symbolizes a microfluidics technology-based miniaturized device that can be scaled down and also able to assimilate various laboratory operations onto a structure that can be fitted on a chip. Various types of inorganic, organic and composite materials are employed to fabricate basic structure LOC devices. Among semiconductors, silicon is employed as the base material for LOC 2 owing to its good chemical stability, structural tunability and medical compatibility. To cut off the large production cost of Si, glass is also employed to build LOC devices having the additional advantage of optical transparency over Si. Among the other materials, lowtemperature cofired ceramic (LTCC) 3 is also used. Various types of organic materials are often employed to fabricate LOC devices such as polymer, paper and hydrogel. Among polymers, two kinds of polymers namely, elastomers and thermoplastic are used. As the performance of the LOC devices largely depends on the fabricated structure, thus, numerous aspects of LOC, along with the required fabrication material was reviewed 4 and the results indicated that the inclusion of nanomaterials will enhance the performance of the LOC 5 irrespective of the material used for structural development. Consequently, various nanomaterials were used in LOC 6 for performance improvement, however, graphene becomes peerless in terms of its inclusion in LOC 7 owing to its novel structure and exotic physio-chemical characteristics. Regarding the fabrication techniques, both top-down and bottom-up methods are employed. Alongside top-down or bottom-up, many sub-processes such as etching, lithography, thermoforming, molding, hot embossing, polymer ablation, polymer casting, bonding are also involved in the final fabrication of the LOC device (Fig 1) . The production procedure and performance of the LOC device are governed by microfluidic technology and the raw material required for the fabrication of such devices should possess some distinct characteristics 9 . In addition, it was also suggested that nanomaterials such as graphene will be the preferred candidate for LOC device fabrication 10 . The honeycomb 4 structure of ultra-lightweight graphene has novel chemical and physical properties 11 likewise extreme optical transparency, outstanding mechanical properties, exceptionally high thermal conductivity, superior electron mobility etc (Fig 2) . The mechanical strength of graphene is very high and it also has the property of pliability. The value of its Young's modulus is 1.0 terapascals with an intrinsic strength of 130 gigapascals, and breaking strength of 42 N/m 13 . The outstanding mechanical strength makes graphene an important material for the fabrication of wearable LOC devices 14 . In addition, graphene possesses excellent electrical conductivity along with a tunable bandgap 15 . Till today, graphene is being considered as the most conductive material 16 are considered as the bottom-up methods for graphene synthesis (Fig 3) . In recent times scientists have devised an advanced graphene synthesis process to curb the cost of production 22 , 23 up to 95.5% 24 and to achieve it they used ultra-low-cost carbon sources 25 . The low production cost of graphene makes it more preferential for the fabrication of inexpensive LOC platforms with superior performance (Fig 4) . The graphene grown by the above-mentioned methods is utilized in various LOC platforms to perform chemical, biological and point of care analysis. However, only the graphene-based analytical LOC devices used for the detection of viruses are reviewed in this paper. Recently, some reviews were published where the use of LOC devices for the detection of only respiratory viruses 27 and the feasibility of using LOC devices for the detection of SARS- CoV-2 28 have been discussed. The graphene-based analytical LOC devices were reported to be employed for the detection of a wide range of viruses' starting from years old Influenza virus to current SARS-CoV-2. Thus, the widest possible range of viruses that are detectable by graphene-based analytical LOC devices is encompassed in this review. Nanoparticles of noble elements are widely used along with graphene for the detection of viruses. Huang et al. 29 prepared a sandwich-type immunosensor for the detection of avian influenza virus H7 (AIV H7) employing graphene coated with silver nanoparticles with a lower limit of detection (LOD) of 1.6 pg/mL. Anik et al. 30 proposed a unique flow-through microfluidic approach for the detection of influenza virus gene employing rGO based transistor exhibiting LOD of 5 pM with high stability and sensitivity. The microfluidic approach was also adopted by Singh et al. 33 to fabricate rGO based electrochemical immunosensor for the detection of H1N1 with LOD of 0.5 PFU mL −1 ( Fig 5) . The cost of the sensor has always been a major issue to make it market-ready. Working in that direction, Joshi et al. 34 GFET was also used by this group 38 for the detection of the influenza virus present in the including biosensor. Chen et al. 39 used this technique to develop a portable GFET biosensor for the detection of the H1N1 virus with a LOD of 1 ng/ml. The fluorometric approach was used by Jeong et al. 40 to fabricate a detection system for influenza virus detection. Multifunctional plasmonic/magnetic graphene was prepared by Lee et al. 41 displayed unique plasmonic and magnetic effects that can be used for the detection of the Influenza virus. The detection technique was highly selective with a lower detection limit of 6.07 pg/ml in human serum. Kinnamon et al. 42 developed a novel screen printed GO-based textile biosensor with high reproducibility and stability. The fabricated biosensor depicted a LOD of 10 ng/mL. Fu et al. 44 Jin et al. 49 They also reported similar biosensing techniques 52 , 53 using rGO-polyamidoamine nanocomposite which showed a better performance with LOD of 0.08 pM for the detection of dengue virus. The fluorometric approach was adopted by Lee et al. 54 and integrated with a loop-mediated isothermal amplification assay to increase the overall sensitivity of the GObased biosensor for the detection of dengue with LOD 2.1 nM. Kamil et al. 55 employed a GO integrated biofunctionalized tapered optical fibre based sensor to detect the dengue virus with a LOD of 1 pM along with the sensitivity of 12.77 nm/nM. Electrochemical impedance spectroscopy was employed by Navakul et al. 56 using GO reinforced polymer to detect the dengue virus with a 0.12 pfu/mL detection limit. Graphene was functionalized with SARS-CoV-2 spike antibody by Seo et al. 57 Liu et al. 68 Jung et al. 72 developed a GO-based immunobiosensor system to detect norovirus employing fluorescence quenching effect between GO and Au nanoparticle. The maximized GO quenching efficiency was obtained up to 85% at 10 5 pfu/mL. Gold nanoparticle combined with graphene plays a significant role in the detection of hepatitis B virus. Huang et al. 73 Basu et al. 83 to develop an rGO based GFET for sensing the hepatitis B virus to achieve a LOD of 50aM. Kim et al. 84 for electrochemical sensing of Hepatitis C virus with a detection limit as low as 160.4 pmol/L. Huang et al. 89 Graphene decorated with nanoparticles was extensively used in determining the presence of norovirus. Lee et al. 94 Gong et al. 102 Photolithography along with plasma-enhanced chemical vapour deposition was employed by The Finally, all the above-mentioned features of the LOC unit are required to be implemented in a cost-effective approach because of the large-scale spreading aspect of a virus outbreak that can even lead to another pandemic scenario without immediate attention (Fig 8) . Graphene-based LOC devices offer the advantages of novel physiochemical properties of graphene along with exotic analytical characteristics of microfluidics namely minimal consumption of target molecules, rapid detection, user-friendliness with high sensitivity. Here, firstly the fabrication process technologies for LOC device has been discussed and thereafter advantages of using graphene in LOC devices have been explored. Later on, the application of graphene-based LOC in detecting a wide spectrum of viruses has been reviewed which ranges from the detection of years old influenza virus to modern SARS-CoV-2. From the economic perspective, graphene depicts a huge potential as recent developments show a steep decrease in the production cost of high-quality graphene. However, still, there are some issues namely toxicity, shelf life and sustainability, which are needed to be addressed with utmost concern. Reproducibility plays a major role in any type of nanodevice and graphene-based LOC comes under it. Moreover, as graphene-based LOCs are mostly meant for point of care related application, thus the process integrity and material reproducibility need an extensive quality check. In summary, till now graphene along with its derivatives has overcome most of the hurdles, thus it can be believed that an eco-friendly, inexpensive, reproducible and sustainable graphene-based LOC platform will pave a new way for the detection of almost all kinds of viruses. Wearable Multifunctional Printed Graphene Sensors Direct Observation of a Widely Tunable Bandgap in Bilayer Graphene Giant Intrinsic Carrier Mobilities in Graphene and Its Bilayer Quantifying the Influence of Graphene Film Nanostructure on the Macroscopic Electrical Conductivity Measuring the Specific Surface Area of Monolayer Graphene Oxide in Water A Highly Efficient Synthetic Process of Graphene Films with Tunable Optical Properties Recent Advances in the Fabrication and Application of Graphene Microfluidic Sensors Precision Synthesis versus Bulk-Scale Fabrication of Graphenes' Machining Induced Transformation of Graphite Flakes to Graphite/Graphene Nanoplatelets Eco-Friendly Production of High Quality Low Cost Graphene and Its Application in Lithium Ion Batteries Novel and Highly Efficient Strategy for the Green Synthesis of Soluble Graphene by Aqueous Polyphenol Extracts of Eucalyptus Bark and Its Applications in High-Performance Supercapacitors Gram-Scale Bottom-up Flash Graphene Synthesis Recent Advances in Graphene-Based Biosensor Technology with Applications in Life Sciences Biosensors for the Detection of Respiratory Viruses: A Review Scalable COVID-19 Detection Enabled by Lab-on-Chip Biosensors Silver Nanoparticles Coated Graphene Electrochemical Sensor for the Ultrasensitive Analysis of Avian Influenza Virus H7 Towards the Electrochemical Diagnostic of Influenza Virus: Development of a Graphene-Au Hybrid Nanocomposite Modified Influenza Virus Biosensor Based on Neuraminidase Activity Dual Immunosensor Based on Methylene Blue-Electroadsorbed Graphene Oxide for Rapid Detection of the Influenza A Virus Antigen A Microfluidic Flow-through Chip Integrated with Reduced Graphene Oxide Transistor for Influenza Virus Gene Detection Label-Free Detection of Influenza Viruses Using a Reduced Graphene Oxide-Based Electrochemical Immunosensor Integrated with a Microfluidic Platform Low Cost Synthesis of Reduced Graphene Oxide Using Biopolymer for Influenza Virus Sensor Graphene Functionalized Field-Effect Transistors for Ultrasensitive Detection of Japanese Encephalitis and Avian Influenza Virus Graphene Field-Effect Transistor for Biosensor Glycan-Functionalized Graphene-FETs toward Selective Detection of Human-Infectious Avian Influenza Virus High-Sensitive and Selective Detection of Human-Infectious Influenza Virus Using Biomimetic Graphene Field-Effect Transistor Wireless Portable Graphene-FET Biosensor for Detecting H 1 N 1 Virus Fluorometric Detection of Influenza Viral RNA Using Graphene Oxide Plasmonic/Magnetic Graphene-Based Magnetofluoro-Immunosensing Platform for Virus Detection Screen Printed Graphene Oxide Textile Biosensor for Applications in Inexpensive and Wearable Point-of-Exposure Detection of Influenza for At-Risk Populations Label-Free Detection of Influenza Viruses Using a Reduced Graphene Oxide-Based Electrochemical Immunosensor Integrated with a Microfluidic Platform Supramolecular Fluorogenic Peptide Sensor Array Based on Graphene Oxide for the Differential Sensing of Ebola Virus Study on Rolling Circle Amplification of Ebola Virus and Fluorescence Detection Based on Graphene Oxide Resonance-Frequency Modulation for Rapid, Point-of-Care Ebola-Glycoprotein Diagnosis with a Graphene-Based Field-Effect Biotransistor A Field Effect Transistor Modified with Reduced Graphene Oxide for Immunodetection of Ebola Virus Field-Effect Transistor Biosensor for Rapid Detection of Ebola Antigen Impedimetric Dengue Biosensor Based on Functionalized Graphene Oxide Wrapped Silica Particles Opto-Electrochemical Functionality of Ru(II)-Reinforced Graphene Oxide Nanosheets for Immunosensing of Dengue Virus Non-Structural 1 Protein Sensitive Surface Plasmon Resonance Performance of Cadmium Sulfide Quantum Dots-Amine Functionalized Graphene Oxide Based Thin Film towards Dengue Virus E-Protein Quantitative and Selective Surface Plasmon Resonance Response Based on a Reduced Graphene Oxide-Polyamidoamine Nanocomposite for Detection of Dengue Virus E-Proteins Sensitive Detection of Dengue Virus Type 2 E-Proteins Signals Using Self-Assembled Monolayers/Reduced Graphene Oxide-PAMAM Dendrimer Thin Film-SPR Optical Sensor Graphene Oxide-Based Molecular Diagnostic Biosensor for Simultaneous Detection of Zika and Dengue Viruses Dengue E Protein Detection Using a Graphene Oxide Integrated Tapered Optical Fiber Sensor A Novel Method for Dengue Virus Detection and Antibody Screening Using a Graphene-Polymer Based Electrochemical Biosensor Rapid Detection of COVID-19 Causative Virus (SARS-CoV-2) in Human Nasopharyngeal Swab Specimens Using Field-Effect Transistor-Based Biosensor Electrical Probing of COVID-19 Spike Protein Receptor Binding Domain via a Graphene Field-Effect Transistor Rapid, Ultrasensitive, and Quantitative Detection of SARS-CoV-2 Using Antisense Oligonucleotides Directed Electrochemical Biosensor Chip Graphene-Based Optical Sensors for the Pre-Vention of SARS-CoV-2 Viral Dissemination Ultrasensitive Supersandwich-Type Electrochemical Sensor for SARS-CoV-2 from the Infected COVID-19 Patients Using a Smartphone Ultra-Sensitive Viral Glycoprotein Detection NanoSystem toward Accurate Tracing SARS-CoV-2 in Biological/Non-Biological Media SARS-CoV-2 RapidPlex: A Graphene-Based Multiplexed Telemedicine Platform for Rapid and Low-Cost COVID-19 Diagnosis and Monitoring Paper-Based Electrochemical Biosensor for Diagnosing COVID-19: Detection of SARS-CoV-2 Antibodies and Antigen Sustainable and Fast Saliva-Based COVID-19 Virus Diagnosis Kit Using a Novel GO-Decorated Au/FBG Sensor Sensing of COVID-19 Antibodies in Seconds via Aerosol Jet Nanoprinted Reduced-Graphene-Oxide-Coated 3D Electrodes Rapid, Ultrasensitive, and Quantitative Detection of SARS-CoV-2 Using Antisense Oligonucleotides Directed Electrochemical Biosensor Chip Micropatterned Reduced Graphene Oxide Based Field-Effect Transistor for Real-Time Virus Detection Graphene-Based Electrochemical Biosensor for Pathogenic Virus Detection Detection of Rota Virus with the Help of Nanomaterial Based Field Effect Transistor (BIO-FET) Accurate Identification and Selective Removal of Rotavirus Using a Plasmonic-Magnetic 3D Graphene Oxide Architecture Graphene Oxide-Based Immunobiosensor for Ultrasensitive Pathogen Detection Disposable Immunoassay for Hepatitis B Surface Antigen Based on a Graphene Paste Electrode Functionalized with Gold Nanoparticles and a Nafion-Cysteine Conjugate Graphene Oxide Wrapped with Gold Nanorods as a Tag in a SERS Based Immunoassay for the Hepatitis B Surface Antigen Gold Nanoparticle-Decorated Reduced-Graphene Oxide Targeting Anti Hepatitis B Virus Core Antigen Real-Time Detection of Hepatitis B Surface Antigen Using a Hybrid Graphene-Gold Nanoparticle Biosensor Design of Aptamer-Based Sensing Platform Using Gold Nanoparticles Functionalized Reduced Graphene Oxide for Ultrasensitive Detection of Hepatitis B Virus', Chemical Papers Label-Free Amperometric Immunosensor Based on Graphene Oxide and Ferrocene-Chitosan Nanocomposites for Detection of Hepatis B Virus Antigen An Electrochemical Immunosensor with Graphene-Oxide-Ferrocene-Based Nanocomposites for Hepatitis B Surface Antigen Detection A Label-Free Electrochemical Platform for the Highly Sensitive Detection of Hepatitis B Virus DNA Using Graphene Quantum Dots A Label-Free Immunosensor for the Sensitive Detection of Hepatitis B e Antigen Based on PdCu Tripod Functionalized Porous Graphene Nanoenzymes Label-Free Biomolecule Detection in Physiological Solutions With Enhanced Sensitivity Using Graphene Nanogrids FET Biosensor Attomolar Sensitivity of FET Biosensor Based on Smooth and Reliable Graphene Nanogrids Deoxyribozyme-Loaded Nano-Graphene Oxide for Simultaneous Sensing and Silencing of the Hepatitis C Virus Gene in Liver Cells Using Silver Nanoparticle and Thiol Graphene Quantum Dots Nanocomposite as a Substratum to Load Antibody for Detection of Hepatitis C Virus Core Antigen: Electrochemical Oxidation of Riboflavin Was Used as Redox Probe An Ultrasensitive and Simple Assay for the Hepatitis C Virus Using a Reduced Graphene Oxide-Assisted Hybridization Chain Reaction Sensitive Electrochemical Detection of Hepatitis C Virus Subtype Based on Nucleotides Assisted Magnetic Reduced Graphene Oxide-Copper Nano-Composite' Ultrasensitive Detection of Hepatitis C Virus DNA Subtypes Based on Cucurbituril and Graphene Oxide Nano-Composite An Ultrasensitive Electrochemical DNA Biosensor Based on Graphene/Au Nanorod/Polythionine for Human Papillomavirus DNA Detection Nucleic Aptamer Modified Porous Reduced Graphene Oxide/MoS2 Based Electrodes for Viral Detection: Application to Human Papillomavirus (HPV) Electrochemical Genosensor Based on Carbon Nanotube/Amine-Ionic Liquid Functionalized Reduced Graphene Oxide Nanoplatform for Detection of Human Papillomavirus (HPV16)-Related Head and Neck Cancer Reduced Graphene Oxide-Based Field Effect Transistors for the Detection of E7 Protein of Human Papillomavirus in Saliva Early-Stage Cervical Cancer Diagnosis Based on an Ultra-Sensitive Electrochemical DNA Nanobiosensor for HPV-18 Detection in Real Samples Binary Nanoparticle Graphene Hybrid Structure-Based Highly Sensitive Biosensing Platform for Norovirus-Like Particle Detection Size-Controlled Preparation of Peroxidase-like Graphene-Gold Nanoparticle Hybrids for the Visible Detection of Norovirus-like Particles Microfluidic Platform Integrated with Graphene-Gold Nano-Composite Aptasensor for One-Step Detection of Norovirus Molybdenum Trioxide Nanocubes Aligned on a Graphene Oxide Substrate for the Detection of Norovirus by Surface-Enhanced Raman Scattering Inkjet-Printed Flexible Biosensor Based on Graphene Field Effect Transistor Aptamer-Based Fluorometric Determination of Norovirus Using a Paper-Based Microfluidic Device Non-Stop Aptasensor Capable of Rapidly Monitoring Norovirus in a Sample Microfluidic Platform Integrated with Graphene-Gold Nano-Composite Aptasensor for One-Step Detection of Norovirus A Sensitive Impedimetric DNA Biosensor for the Determination of the HIV Gene Based on Electrochemically Reduced Graphene Oxide A Sensitive Impedimetric DNA Biosensor for the Determination of the HIV Gene Based on Graphene-Nafion Composite Film Sensitive Electrochemical DNA Sensor for the Detection of HIV Based on a Polyaniline/Graphene Nanocomposite A T7exonuclease-Assisted Target Recycling Amplification with Graphene Oxide Acting as the Signal Amplifier for Fluorescence Polarization Detection of Human Immunodeficiency Virus (HIV) DNA' Large-Scale Graphene Micropattern Nano-Biohybrids: High-Performance Transducers for FET-Type Flexible Fluidic HIV Immunoassays A Smart Nanosensor for the Detection of Human Immunodeficiency Virus and Associated Cardiovascular and Arthritis Diseases Using Functionalized Graphene-Based Transistors Attomolar Detection of Virus by Liquid Coplanar-Gate Graphene Transistor on Plastic Scalable Graphene Aptasensors for Drug Quantification Boron and Nitrogen Co-Doped Single-Layered Graphene Quantum Dots: A High-Affinity Platform for Visualizing the Dynamic Invasion of HIV DNA into Living Cells through Fluorescence Resonance Energy Transfer Graphene Oxide-Based Biosensing Platform for Rapid and Sensitive Detection of HIV-1 Protease Graphene Oxide-Polycarbonate Track-Etched Nanosieve Platform for Sensitive Detection of Human Immunodeficiency Virus Envelope Glycoprotein Ultrasensitive Electrochemical Biosensor for HIV Gene Detection Based on Graphene Stabilized Gold Nanoclusters with Exonuclease Amplification Novel Graphene-Based Biosensor for Early Detection of Zika Virus Infection Electrochemical Detection of Zika Virus in Biological Samples: A Step for Diagnosis Point-of-Care Impedimetric Zika and Dengue Biosensor Based on Functionalized Graphene Oxide Wrapped Silica Particles An Impedimetric Immunosensor for Determination of Porcine Epidemic Diarrhea Virus Based on the Nanocomposite Consisting of Molybdenum Disulfide/Reduced Graphene Oxide Decorated with Gold Nanoparticles Electrical Pulse-Induced Electrochemical Biosensor for Hepatitis E Virus Detection Rapid and Fully Automated Bacterial Pathogen Detection on a Centrifugal-Microfluidic LabDisk Using Highly Sensitive Nested PCR with Integrated Sample Preparation A Lab-on-a-Chip System with Integrated Sample Preparation and Loop-Mediated Isothermal Amplification for Rapid and Quantitative Detection of Salmonella Spp. in Food Samples A Rapid, Low-Cost, and Microfluidic Chip-Based System for Parallel Identification of Multiple Pathogens Related to Clinical Pneumonia Development of a Self-Priming PDMS/Paper Hybrid Microfluidic Chip Using Mixed-Dye-Loaded Loop-Mediated Isothermal Amplification Assay for Multiplex Foodborne Pathogens Detection An Integrated Temporary Negative Pressure Assisted Microfluidic Chip for DNA Isolation and Digital PCR Detection Digital PCR-An Emerging Technology with Broad Applications in Microbiology Printable QR Code Paper Microfluidic Colorimetric Assay for Screening Volatile Biomarkers Opportunities and Challenges for Biosensors and Nanoscale Analytical Tools for Pandemics: COVID-19