Urgent Needs in Surveillance and Diagnostics. Zika Firebird Biomolecular Sciences LLC Florida Medical Entomology Laboratory, the University of Florida Steven A. Benner Barry W. Alto ABSTRACT The potential hazards of the Zika virus to both American citizenry and citizens of the world need no discussion. We are still learning the distribution and impact of the virus on human development in utero, as well as its distribution in patient saliva, sexual fluids, and urine. With several dozen cases in the US at the time of this writing, most obtained overseas, the extramural research programs of public health agencies are being asked to seek better public health surveillance technologies and immediate human diagnostics tools for Zika. For example, in Brazil, the immediate need is a test that to distinguish between dengue, chikungunya, and Zika in public clinics; at this time, this distinction is made based on interrogation of patients with respect to the timing of the appearance of various symptoms. The work proposed here will deliver those technologies and tools for Zika. However, this work will provide more. Today, the NIAID, CDC, and other public health units responsible for managing infectious disease outbreaks move from disease to disease, with each new outbreak and news cycle. This is inefficient, at best. Efficient public health services in an age of emerg- ing diseases need a robust and reproducible technology platform that is sufficiently flexible to sur- vey for any emerging infectious diseases without crisis. The work proposed here offers such a platform as part of its ability to survey for Zika, and distinguish it from other arboviruses, based on reagent innovations drawn from the field of ?synthetic biology?, including artificially expanded genetic infor- mation systems (AEGIS), self-avoiding molecular recognition systems (SAMRS), biversal nucleotides, transliteration, and evolution-based arbovirus sequence analysis, in a collaboration between the Flor- ida Medical Entomology Laboratory (FMEL), which has live lab-infected mosquitoes with Zika, dengue, and chikungunya, and Firebird, a serial innovator in synthetic biology and diagnostics technology. This R21 project will first add Zika to an already established kit that detects 22 other arboviruses, common and exotic.13 We will benchmark that expanded kit, delivering it to beta testers in FMEL, where multiplexed readout is done using a Luminex instrument. This independent testing guarantees scientific rigor, and will be done by Month 12. In Year 2, we will seek a simpler system, focusing the multiplex on geography-specific target sets, adding biversals to manage sequence evolution of the RNA viruses, replacing Luminex in 10x target assays by a readout based on a 4x4 micro array, and going as far as possible towards a 10x multiplex system that can assay ~ 10 mosquitoes in the field. Towards these ends, we will meet the following specific aims: The outcome of this project will include new Zika science, as well as kits that will allow public health services to enjoy the advantages of these reagent innovations in managing the Zika outbreak, just as those advantages are now enjoyed in many products, including FDA-approved products, for human diagnostics.

Public Health Relevance

Meeting Urgent Needs in Surveillance and Diagnostics. Zika Firebird Biomolecular Sciences LLC Florida Medical Entomology Laboratory, the University of Florida Steven A. Benner Barry W. Alto NARRATIVE The outbreak of Zika virus in Latin America, the presence of the virus in the US in patients returning from outbreak regions, and the potential for an outbreak of Zika in the United States, together with the possibility that Zika disrupts the normal development of babies in utero, has motivated President Obama to allocate an additional $1.8 billion to research and development to manage public health concerns. The premise of this research is that a low-cost, multipurpose kit that allows public health officials to identify Zika in trapped mosquitoes will help in that management, a premise that is not disputed in the public health community. The impact of these kits will be still greater if it is set up to, in the next phase of research, transition into a low-cost kits that can detect Zika virus in saliva and urine at points of sampling in low resource environments. This R21 research program will have further impact by familiarizing the community with the potential of synthetic biology to address public health concerns and, we hope, to move federal activities related to infectious disease from a 'disease of the month' response model (SARS, chikungunya, Valley fever, Ebola, MERS, dengue, and now Zika), where each new outbreak and news cycle creates a crisis, entirely unnecessary if synthetic biology innovations were to be moved into public health management, where a single technology platform gives public health officials a kit that has routine use, but also is flexible enough to accommodate new and exotic pathogens as they emerge.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Exploratory/Developmental Grants (R21)
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Special Emphasis Panel (ZRG1)
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Challberg, Mark D
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Firebird Biomolecular Sciences, LLC
United States
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Yaren, Ozlem; Alto, Barry W; Bradley, Kevin M et al. (2018) Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1. J Vis Exp :
Glushakova, Lyudmyla G; Alto, Barry W; Kim, Myong-Sang et al. (2018) Optimization of cationic (Q)-paper for detection of arboviruses in infected mosquitoes. J Virol Methods 261:71-79
Yaren, Ozlem; Alto, Barry W; Gangodkar, Priyanka V et al. (2017) Point of sampling detection of Zika virus within a multiplexed kit capable of detecting dengue and chikungunya. BMC Infect Dis 17:293