Ultrasensitive, Rapid, Amplification-Free RNA Virus Detection Using Nanodimer-Based Nucleic Acid Target Sequence Recognition

Ribonucleic acid (RNA) viruses are a major threat to public health and global economy. Numerous human diseases, from the common colds to life-threatening hemorrhagic fevers are caused by RNA viruses. In the past decades, RNA viruses such as human immune deficiency virus, Ebola virus, dengue virus, and most recently coronavirus (COVID-19) impose significant burdens on global health and global economy. Rapid and high-sensitivity detection of the presence of RNA viruses in human specimens and the environment is crucial to track the rate of viral contamination, prevent fast spreading, and establish preventive measures and timely administration of treatments. The popular polymerase chain reaction (PCR)-based virus detection takes lengthy turn-around time and requires specialized laboratory instruments and trained personnel. This project aims to create a novel biosensing microsystem that can provide ultra-sensitive, rapid and reliable detection of RNA viruses with no need of PCR amplification. The project will result in a portable, rapid, economical RNA virus detection method for point-of-care-testing, pandemic prevention, anti-bioterrorism, and environmental monitoring. Furthermore, this cross-disciplinary research will support the development of a diverse cohort of graduate, undergraduate students at the University of Akron. The research results will also support several graduate-level courses on microsystems and biomaterials at the University of Akron. The technical scope of the project are divided into multiple tasks: 1) create a surface acoustic pre-filtering chip that will rapidly remove microscale impurities in continuous flow, which would otherwise cause clogging of the sensing channels, 2) research a novel nanoparticle focusing method based on unique electro-diffusio-phoresis effect, which can significantly increases the virion concentration and thus output signals thousands of times within seconds, 3) research a virus detection chip based on an innovative nanodimer dissociation assay, which in principle, can detect viruses with single virus resolution, 4) research unique signal multiplexing applied on a resistive pulse sensor array, which will enable high throughput, digital detection of RNA viruses, and ultimately 5) integrate the three chips into a microsystem and demonstrate its utility for rapid detection of ultra-low abundance RNA viruses using an inactivated model virus. The research will advance the field of virus detection with ultra-high resolution and digitization capability, which are difficult to achieve using current state-of-the-art methods. It will also generate numerous new knowledges and innovations for the development of the next generation high-throughput, onsite biosensing systems in general. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.