Diagnostic reagents and assays to detect biodefense pathogens are critical needs for public safety. Two key components for a successful molecular diagnostic assay are a sensor component that binds directly to the targeted organism, or to a product secreted by the organism, and a signal domain that indicates, with great sensitivity, when the sensor has bound the target molecule. Over the past 18 months we have implemented proven technologies: phage-display, covalent protein-DNA linkage, and real-time PCR, along with the results from our antigen discovery research, to begin to create a powerful diagnostic assay to detect the presence of Francisella tularensis (Ft) in biological and environmental samples as well as immune responses directed against Ft. The protein-DNA chimeras central to these assays are called tadpoles, which are capable of achieving a much greater level of sensitivity (~106-fold greater) compared to analogous enzyme-linked immunosorbent assays (ELISAs). Furthermore these diagnostics are able to identify target molecules over a wide dynamic range of concentrations. In this proposal, we will multiplex the tadpole assay to include additional fever and diarrheal agents as outlined in the overall WRCE diagnostic theme plan. Ft, Rift Valley fever virus, and Cryptosporidium parvum will serve as initial, comparative controls for the platform WRCE diagnostic approaches, and the agent list will be expanded in later years according to the WRCE plan. Furthermore, we will seek to integrate elemental technologies from other platforms being developed in the WRCE diagnostic group, such as lateral flow microfluidics, to expand the utility of tadpole diagnostics as they are further multiplexed for the simultaneous detection of numerous agents and are adapted for point-of-care usage in addition to their use in the reference laboratory.

Public Health Relevance

Diagnostics that can detect minute quantities of a biodefense pathogen organism and/or that can detect whether a person has been infected by such an organism early in the course of infection are greatly needed. We will use cutting-edge strategies to create diagnostic reagents and assays that can identify tiny amounts of multiple biodefense pathogens in complex samples to a high degree of sensitivity. The strategies applied here can easily be adapted to create similar reagents for any biodefense or infectious disease concern.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Specialized Center--Cooperative Agreements (U54)
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Special Emphasis Panel (ZAI1-DDS-M)
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University of Texas Medical Br Galveston
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Paterson, Andrew S; Raja, Balakrishnan; Garvey, Gavin et al. (2014) Persistent luminescence strontium aluminate nanoparticles as reporters in lateral flow assays. Anal Chem 86:9481-8
Santiago, Felix W; Covaleda, Lina M; Sanchez-Aparicio, Maria T et al. (2014) Hijacking of RIG-I signaling proteins into virus-induced cytoplasmic structures correlates with the inhibition of type I interferon responses. J Virol 88:4572-85
Pflughoeft, Kathryn J; Swick, Michelle C; Engler, David A et al. (2014) Modulation of the Bacillus anthracis secretome by the immune inhibitor A1 protease. J Bacteriol 196:424-35
Lavinder, Jason J; Wine, Yariv; Giesecke, Claudia et al. (2014) Identification and characterization of the constituent human serum antibodies elicited by vaccination. Proc Natl Acad Sci U S A 111:2259-64
Valbuena, Gustavo; Halliday, Hailey; Borisevich, Viktoriya et al. (2014) A human lung xenograft mouse model of Nipah virus infection. PLoS Pathog 10:e1004063
Nieves, Wildaliz; Petersen, Hailey; Judy, Barbara M et al. (2014) A Burkholderia pseudomallei outer membrane vesicle vaccine provides protection against lethal sepsis. Clin Vaccine Immunol 21:747-54
Gardner, Christina L; Hritz, Jozef; Sun, Chengqun et al. (2014) Deliberate attenuation of chikungunya virus by adaptation to heparan sulfate-dependent infectivity: a model for rational arboviral vaccine design. PLoS Negl Trop Dis 8:e2719
Litvinov, Julia; Hagström, Anna E V; Lopez, Yubitza et al. (2014) Ultrasensitive immuno-detection using viral nanoparticles with modular assembly using genetically-directed biotinylation. Biotechnol Lett 36:1863-8
Caro-Gomez, Erika; Gazi, Michal; Goez, Yenny et al. (2014) Discovery of novel cross-protective Rickettsia prowazekii T-cell antigens using a combined reverse vaccinology and in vivo screening approach. Vaccine 32:4968-76
Georgiou, George; Ippolito, Gregory C; Beausang, John et al. (2014) The promise and challenge of high-throughput sequencing of the antibody repertoire. Nat Biotechnol 32:158-68

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