Treatments and hospitalizations due to respiratory infections incur costs exceeding $40 billion dollars yearly in the US alone. The impact of respiratory disease on human health is even greater worldwide and the potential of a global epidemic involving respiratory viruses is high. Currently, the most sensitive and comprehensive assays fail to detect a viral pathogen in at least half of clinical samples from individuals with clear respiratory symptoms. Most of these assays are expensive and not intended for point-of-care use. In this Phase I application, we propose to combine two innovative platform technologies: our molecular approach, using a unique pool of consensus-degenerate hybrid oligonucleotide primers (CODEHOPs) to identify novel members of pathogen families by PCR amplification, with a fully integrated, reagent containing microfluidics cartridge capable of rapid and accurate testing that we currently have in commercial development. This will provide a first ever nucleic acid assay based, cost-effective and robust prototype molecular platform suitable for point-of- care diagnosis and surveillance. The intended commercial use of this unique combination of technologies is as a screening method to detect viruses in specimens for which known viruses have not been detected and for global surveillance and epidemiological studies of emerging infections. Our objective is to demonstrate the feasibility of this diagnostic tool by targeting two prominent respiratory virus families, adenoviruses and parainfluenzaviruses, that constitute a significant public health burden.
The specific aims are (1) to develop and validate real-time CODEHOP PCR assays for broad-based detection of members of the adenovirus and paramyxovirus families (2) to transfer these assays to microfluidics cartridges enabling fast, inexpensive, and easy-to-use detection of respiratory viruses at point-of-care;and (3) to verify CODEHOP on-cartridge detection for ten clinical isolates from each family. Achieving these aims would support a Phase II application that would focus on the development of a novel viral strain-specific solid-phase (VISP) test based on the CODEHOP amplicons for each virus family to rapidly distinguish viral species amplified in the PCR reaction, and on the integration of the assay into a microfluidics-based detection system. Phase II development would also broaden the targeted panel of respiratory virus families to include all viruses causing respiratory disease. The final product will comprise disposable, single-use microfluidics cartridges, heat-stable agents and a portable instrument (beta system will be available during Phase II development) to operate the fully integrated cartridge and provide the read-out. This cartridge-based system will be capable of detecting viral respiratory pathogens at point-of-care even if the causative pathogen is unknown. We expect that the development of such a rapid and easy-to-use diagnostic product would provide physicians in small clinics and hospitals with a much needed tool for comprehensive testing of respiratory diseases at point-of-care. This could have a large impact on our surveillance capacities as well as prevention and treatment strategies and ultimately improve human health.
Widespread nucleic acid assay testing of patients with respiratory symptoms in small hospitals, clinics and physician's offices is not possible today. We propose the development of a comprehensive and affordable point-of-care microfluidics-based system that is uniquely able to detect viral respiratory pathogens even if the causative pathogen was previously unknown. This system is expected to yield higher rates of diagnosis and allow detection of emerging epidemic threats. The system will have the potential to improve patient outcomes by immediate tailoring of the therapy to the virus, reducing antibiotic use and unnecessary testing, and decreasing the number and times of hospitalization. It will provide data necessary to manage outbreaks and minimize community impact. Since respiratory illnesses are so prominent all this clearly has the potential to reduce health-care costs enormously and improve public health significantly.
