This proposal describes the design, construction, and testing of a rapid multiplexed matrix code diagnostic for select biodefense and emerging pathogens. The threat of biological attacks underscores a critical need for rapid point-of care diagnostics that identify specific pathogens in infected persons. Infected or at-risk individuals may be far from critical-care facilities;therefore, diagnostic tests must be easily transportable, stable in storage, simple to use, and easy to interpret. Lateral flow tests (also called immunochromatographic tests or strip tests) are a well-established diagnostic platform because of their simplicity, rapid readout, and low cost qualitative/semi-quantitative detection of antibodies or antigens. Nonetheless, the lack of available point-of-care diagnostics for infectious diseases represents vulnerability in biodefense preparedness as well as a gap in serving public health needs. We propose novel diagnostic device designs, including quick response (QR) two-dimensional matrix-coded readouts, along with attention to chemical coupling approaches, to increase the sensitivity and broaden the multiplex capabilities of lateral flow devices. Effective public health responses also require knowledge of disease distribution and movement based on diagnostic data. Therefore, our proposed Multiplexed Matrix Diagnostic (MMDx) device will be machine-readable, representing a field diagnostic with integrated capability for real time epidemiology using mobile phone technologies and geographic positioning. We have assembled an interdisciplinary team of scientists, engineers, and medical doctors who understand the biology of, and immune responses to infectious agents, the physical chemistry of detection methods, the fabrication methods required to produce diagnostic devices, the technology for providing real-time responsive epidemiology, and the processes required for validating a diagnostic device under field conditions. Our goals for the exploratory/pilot (R21) phase are to develop a prototype two-dimensional matrix barcode-based, machine-readable diagnostic to detect pathogens in blood or serum. The prototype will offer 1) direct identification of the four different dengue virus serotypes by detecting circulating viral protein NS1, as well as 2) indirect Ebola virus and dengue virus identification by detecting circulating antibodies against the viral proteins. A field-based feasibility test archival patient serum samples collected from dengue virus-infected individuals will provide initial validation of the specificity and sensitivit of the prototype and enable power calculations to determine numbers of required patients prior to the R33 developmental phase. For the Developmental Phase (R33), our goals will be to extend the MMDx device multiplex capabilities for detecting additional biodefense agent infections in a point-of-care analysis. The full potential of the MMDx diagnostic and real-time epidemiology will be evaluated in a one-month hospital-based field test in Bucaramanga, Colombia, which has a high incidence of dengue virus infections.
A bioterror attack could introduce an unknown pathogen into an area, causing disease without a familiar diagnosis. Protecting and treating military and civilian personnel requires rapid disease diagnosis. This proposal describes the design, construction, and testing of a novel diagnostic for concurrent detection of multiple pathogens in a setting where no specialized equipment or expertise is required.
|Myhrvold, Cameron; Freije, Catherine A; Gootenberg, Jonathan S et al. (2018) Field-deployable viral diagnostics using CRISPR-Cas13. Science 360:444-448|
|Muffat, Julien; Li, Yun; Omer, Attya et al. (2018) Human induced pluripotent stem cell-derived glial cells and neural progenitors display divergent responses to Zika and dengue infections. Proc Natl Acad Sci U S A 115:7117-7122|
|Tam, Justina O; de Puig, Helena; Yen, Chun-Wan et al. (2017) A comparison of nanoparticle-antibody conjugation strategies in sandwich immunoassays. J Immunoassay Immunochem 38:355-377|
|de Puig, Helena; Bosch, Irene; Gehrke, Lee et al. (2017) Challenges of the Nano-Bio Interface in Lateral Flow and Dipstick Immunoassays. Trends Biotechnol 35:1169-1180|
|Metsky, Hayden C; Matranga, Christian B; Wohl, Shirlee et al. (2017) Zika virus evolution and spread in the Americas. Nature 546:411-415|
|de Puig, Helena; Bosch, Irene; Carré-Camps, Marc et al. (2017) Effect of the Protein Corona on Antibody-Antigen Binding in Nanoparticle Sandwich Immunoassays. Bioconjug Chem 28:230-238|
|Bosch, Irene; de Puig, Helena; Hiley, Megan et al. (2017) Rapid antigen tests for dengue virus serotypes and Zika virus in patient serum. Sci Transl Med 9:|
|Durbin, Ann Fiegen; Wang, Chen; Marcotrigiano, Joseph et al. (2016) RNAs Containing Modified Nucleotides Fail To Trigger RIG-I Conformational Changes for Innate Immune Signaling. MBio 7:|
|Yen, Chun-Wan; de Puig, Helena; Tam, Justina O et al. (2015) Multicolored silver nanoparticles for multiplexed disease diagnostics: distinguishing dengue, yellow fever, and Ebola viruses. Lab Chip 15:1638-41|
|de Puig, Helena; Tam, Justina O; Yen, Chun-Wan et al. (2015) Extinction Coefficient of Gold Nanostars. J Phys Chem C Nanomater Interfaces 119:17408-17415|