Arthropod-borne viruses (arboviruses) comprise many of the most important ?emerging pathogens? due to their geographic spread and their increasing impact on vulnerable human populations. There is urgent need for easy-to- operate and rapidly deployable diagnostic tools that can handle blood samples in a closed sample-to-answer manner. Here, we propose to develop a novel diagnostic technology that can detect viral antigens in an inexpensive, ultrasensitive, specific, and multiplexed manner. We will develop our novel approach into standalone tool with a detection capability at attomolar sensitivities (comparable to nucleic acid amplification tests) to diagnose arboviral infections with minimal user interference. The integrated diagnostic platform will utilize a novel surrogate approach, microfluidic integration, and a multiplexed detection scheme with the capacity to distinguish arboviral infections. The system will be designed to initiate diagnosis from serum/plasma/blood and provide a sample-to-answer diagnostic within less than 35 minutes using less than 100 L blood samples at a cost of $2 per test. Collaborative work proposed for this NIH/NIAID R01 Grant involves integration of nanophotonic engineering (Yanik Group), molecular virology (Pinsky Group), and infectious diseases epidemiology (LaBeaud Group) to build and field-test our novel point-of-care viral diagnostic platform with Windward Islands Research and Education Foundation (WINDREF) and St. George?s University teams (Macpherson, Waechter and Noel Groups). Preliminary validation tests with patient samples will be initially performed at Stanford Medical Facility in collaboration with LaBeaud and Pinsky groups. Subsequently, three prototypes will be transferred to Grenada for field-testing initially at central laboratories then to resource-poor settings in small towns. Yanik group will provide the necessary expertise for integration of molecular and nanoengineering components and demonstration of a practical prototype as well as evaluating the application of prototype(s) developed under this proposal with patient samples (LaBeaud and Pinsky Groups). System will be iteratively optimized and a rugged platform suitable for field settings will be developed.
The goal of this proposal is to develop a rapid, sensitive, low-cost, and easy-to-use diagnostic technology for diagnostics of arboviral infections in point-of-care settings. In our preliminary experiments with serum samples, we demonstrated that our novel approach is 10,000 fold more sensitive than the most sensitive ELISA tests. Our goal is to develop our novel approach into a standalone diagnostic unit, validate it with PCR measurements, and field-test in Grenada. The proposed studies will take advantage of the considerable resources and expertise at University of California, Santa Cruz (UCSC) and Stanford University, the established febrile human disease surveillance and archived serum banks from Kenya, the established human disease surveillance in Grenada, as well as industry expertise in development and production of antibodies specific to non-structural 1 protein (NS1), recombinant NS1 proteins and anti-IgM antibodies.