Modern isolation and separation techniques for extracellular vesicles.

Infectious diseases account for more than 20% of global mortality and viruses are responsible for about one-third of these deaths. Highly infectious viral diseases such as severe acute respiratory (SARS), Middle East respiratory syndrome (MERS) and coronavirus disease (COVID-19) are emerging more frequently and their worldwide spread poses a serious threat to human health and the global economy. The current COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). As of 27 July 2020, SARS-CoV-2 has infected over 16 million people and led to the death of more than 652,434 individuals as on 27 July 2020 while also causing significant economic losses. To date, there are no vaccines or specific antiviral drugs to prevent or treat COVID-19. Hence, it is necessary to accelerate the development of antiviral drugs and vaccines to help mitigate this pandemic. Non-Conventional antiviral agents must also be considered and exploited. In this regard, nanoparticles can be used as antiviral agents for the treatment of various viral infections. The use of nanoparticles provides an interesting opportunity for the development of novel antiviral therapies with a low probability of developing drug resistance compared to conventional chemical-based antiviral therapies. In this review, we first discuss viral mechanisms of entry into host cells and then we detail the major and important types of nanomaterials that could be used as antiviral agents. These nanomaterials include silver, gold, quantum dots, organic nanoparticles, liposomes, dendrimers and polymers. Further, we consider antiviral mechanisms, the effects of nanoparticles on coronaviruses and therapeutic approaches of nanoparticles. Finally, we provide our perspective on the future of nanoparticles in the fight against viral infections.

Antiviral Potential of Nanoparticles-Can Nanoparticles Fight Against Coronaviruses?

Many problems exist within the myriad of currently employed screening and diagnostic methods. Further, an incredibly wide variety of procedures are used to identify an even greater number of diseases which exist in the world. There is a definite unmet clinical need to improve diagnostic capabilities of these procedures, including improving test sensitivity and specificity, objectivity and definitiveness, and reducing cost and invasiveness of the test, with an interest in replacing multiple diagnostic methods with one powerful tool. There has been a recent surge in the literature which focuses on utilizing Raman spectroscopy in combination with machine learning analyses to improve diagnostic measures for identifying an assortment of diseases, including cancers, viral and bacterial infections, neurodegenerative and autoimmune disorders, and more. This review highlights the work accomplished since 2018 which focuses on using Raman spectroscopy and machine learning to address the need for better screening and medical diagnostics in all areas of disease. A critical evaluation considers both the benefits and obstacles of utilizing the method for universal diagnostics. It is clear based on the evidence provided herein Raman spectroscopy in combination with machine learning provides the first glimmer of hope for the development of an accurate, inexpensive, fast, and non-invasive method for universal medical diagnostics.

Towards development of a novel universal medical diagnostic method: Raman spectroscopy and machine learning

Since late December 2019, the coronavirus pandemic (COVID-19; previously known as 2019-nCoV) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been surging rapidly around the world. With more than 1,700,000 confirmed cases, the world faces an unprecedented economic, social, and health impact. The early, rapid, sensitive, and accurate diagnosis of viral infection provides rapid responses for public health surveillance, prevention, and control of contagious diffusion. More than 30% of the confirmed cases are asymptomatic, and the high false-negative rate (FNR) of a single assay requires the development of novel diagnostic techniques, combinative approaches, sampling from different locations, and consecutive detection. The recurrence of discharged patients indicates the need for long-term monitoring and tracking. Diagnostic and therapeutic methods are evolving with a deeper understanding of virus pathology and the potential for relapse. In this Review, a comprehensive summary and comparison of different SARS-CoV-2 diagnostic methods are provided for researchers and clinicians to develop appropriate strategies for the timely and effective detection of SARS-CoV-2. The survey of current biosensors and diagnostic devices for viral nucleic acids, proteins, and particles and chest tomography will provide insight into the development of novel perspective techniques for the diagnosis of COVID-19.

Current and Perspective Diagnostic Techniques for COVID-19.

The growing demand for reliable and robust methodology in bio-chemical sensing calls for the continuous advancement of sensor technologies. Over the last two decades, surface-enhanced Raman spectroscopy (SERS) has emerged as one of the most promising analytical techniques for sensitive and trace analysis or detection in biomedical and agri-food applications. SERS overcomes the inherent sensitivity limitation associated with Raman spectroscopy, which provides vibrational "fingerprint" spectra of molecules that makes it unique and versatile among other spectroscopy techniques. This paper comprehensively reviews the recent advancements of SERS for biomedical, food and agricultural applications over the last 6 years, and we envision that, in the near future, some of these platforms have the potential to be translated as a point-of-care and rapid sensor for real-life end-user applications. The merits and limitations of various SERS sensor designs are analysed and discussed based on critical features such as sensitivity, specificity, usability, repeatability and reproducibility. We conclude by highlighting the opportunities and challenges in the field while stressing the technological gaps to be addressed in realizing commercially viable point-of-care SERS sensors for practical biomedical and agri-food technological applications.

Towards a point-of-care SERS sensor for biomedical and agri-food analysis applications: a review of recent advancements

Emerging and reemerging viruses are responsible for a number of recent epidemic outbreaks. A crucial step in predicting and controlling outbreaks is the timely and accurate characterization of emerging virus strains. We present a portable microfluidic platform containing carbon nanotube arrays with differential filtration porosity for the rapid enrichment and optical identification of viruses. Different emerging strains (or unknown viruses) can be enriched and identified in real time through a multivirus capture component in conjunction with surface-enhanced Raman spectroscopy. More importantly, after viral capture and detection on a chip, viruses remain viable and get purified in a microdevice that permits subsequent in-depth characterizations by various conventional methods. We validated this platform using different subtypes of avian influenza A viruses and human samples with respiratory infections. This technology successfully enriched rhinovirus, influenza virus, and parainfluenza viruses, and maintained the stoichiometric viral proportions when the samples contained more than one type of virus, thus emulating coinfection. Viral capture and detection took only a few minutes with a 70-fold enrichment enhancement; detection could be achieved with as little as 102 EID50/mL (50% egg infective dose per microliter), with a virus specificity of 90%. After enrichment using the device, we demonstrated by sequencing that the abundance of viral-specific reads significantly increased from 4.1 to 31.8% for parainfluenza and from 0.08 to 0.44% for influenza virus. This enrichment method coupled to Raman virus identification constitutes an innovative system that could be used to quickly track and monitor viral outbreaks in real time.

https://www.pnas.org/content/pnas/117/2/895.full.pdf

A rapid and label-free platform for virus capture and identification from clinical samples

Recent discoveries reveal that extracellular vesicles (EVs) play an important role in transmitting signals. Although this emerging transcellular pathway enables a better understanding of neural communication, the lack of techniques for effectively isolating EVs impedes their studies. Herein, we report an emergent high-throughput platform consisting of three-dimensional carbon nanotube arrays that rapidly capture different EVs based on their sizes, without any labels. More importantly, this label-free capture maintains the integrity of the EVs when they are excreted from a host cell, thus allowing comprehensive downstream analyses using conventional approaches. To study neural communication, we developed a stamping technique to construct a gradient of nanotube herringbone arrays and integrated them into a microdevice that allowed us processing of a wide range of sample volumes, microliters to milliliters, in several minutes through a syringe via manual hand pushing and without any sample preparation. This microdevice successfully captured and separated EVs excreted from glial cells into subgroups according to their sizes. During capture, this technology preserved the structural integrity and originality of the EVs that enabled us to monitor and follow internalization of EVs of different sizes by neurons and cells. As a proof of concept, our results showed that smaller EVs (∼80 nm in diameter) have a higher uptake efficiency compared to larger EVs (∼300 nm in diameter). In addition, after being internalized, small EVs could enter endoplasmic reticulum and Golgi but not the largest ones. Our platform significantly shortens sample preparation, allows the profiling of the different EVs based on their size, and facilitates the understanding of extracellular communication. Thus, it leads to early diagnostics and the development of novel therapeutics for neurological diseases.

Rapid Size-Based Isolation of Extracellular Vesicles by Three-Dimensional Carbon Nanotube Arrays

The reach and the rate of spread of viral infectious diseases have vastly increased owing to changes in various environmental, biological, socioeconomic, and political factors as well as expansion of international travel and trade. Consequently, the need for more rapid outbreak detection and efficient monitoring of the spread of the infection to minimize the burden on global health and the economy has also become of paramount importance. Rapid isolation, identification, and full characterization of viruses from field samples are critical in the prevention of the epidemics. In this regard, nanotechnology has emerged as a significant platform for human health care and point-of-care diagnosis of interactions in the cellular and subcellular scales. This chapter presents a review of the exploration of low-dimensional materials, such as carbon nanotubes and transition metal dichalcogenides in developing lab-on-a-chip system applicable in virology as a point-of-care device for size-based virus capture and enrichment for any virus. Future prospects of these materials in epidemiology are also discussed.

Venkataraman Swaminathan

Papers

A rapid and label-free platform for virus capture and identification from clinical samples

Rapid Size-Based Isolation of Extracellular Vesicles by Three-Dimensional Carbon Nanotube Arrays

The application of low-dimensional materials in virology and in the study of living organisms