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Title:Point of care platforms for protein and nucleic acid detection
Author(s):Berger, Jacob
Director of Research:Bashir, Rashid
Doctoral Committee Chair(s):Bashir, Rashid
Doctoral Committee Member(s):King, William P.; Cunningham, Brian; Smith, Andrew
Department / Program:Bioengineering
Discipline:Bioengineering
Degree Granting Institution:University of Illinois at Urbana-Champaign
Degree:Ph.D.
Genre:Dissertation
Subject(s):Point-of-care, microfluidics, assay development
Abstract:Microfluidic systems can allow for the processing of a small volume of sample resulting in a portable and disposable means for rapid quantification for biomedical needs. These systems can detect pathogens, blood and serum-based biomarkers, and identify known disease markers. Many diagnostic systems developed to quantify cellular and proteomic take several hours and require sample purification before producing results. These delays in diagnostic capabilities greatly hinder time-sensitive results for preventing illness progression, where time-sensitive and vital diagnostic information for the doctor and healthcare professionals guide and determine treatment. For example, sepsis is a disease that would greatly benefit from a rapid stratification tool for disease monitoring. Sepsis, a life-threatening organ dysfunction caused by a dysregulated host response, leads the U.S in both mortality rate and cost of treatment. Sepsis treatment protocols currently rely on broad and non-specific parameters like heart and respiration rate, and temperature; however, studies show that biomarkers Interlukin-6 (IL-6) and Procalcitonin (PCT) correlate to sepsis progression and response to treatment. Prior work also suggests that using multi-parameter predictive analytics with biomarkers and clinical information can inform treatment to improve outcomes. A point-of-care (POC) platform that provides information for multiple biomarkers can aid in the diagnosis and prognosis of potentially septic patients. This thesis reports a microfluidic POC system that correlates microbead capture to IL-6 and PCT concentrations using impedance cytometry, microbead immunoassays, and biotin-streptavidin binding. A multiplexed microbead immunoassay is developed and validated first for the detection of IL-6. The assay principles are further used to develop microbead based detection for PCT. The final platform demonstrates the simultaneous detection of both IL-6 and PCT from human plasma samples. Aside from utilizing microfluidics for diagnostics of disease biomarkers at the point of care, the inability to diagnose infections in both low resource settings have produced many economic and social problems. Many disease outbreaks that occur from community transmission and recent advancements in globalization require diagnostic technologies than can both serve as a monitoring system and respond to exponential rates of transmission. When developing POC diagnostic platforms in a research setting, many the developed microfluidics platform are often fabricated from a combination of silicon, silicone, and glass. However, to translate these POC platforms from the laboratory to the clinic, the manufacturing of these devices must be reengineered to have the same properties and diagnostic capabilities when manufactured from a from a polymer typically made with an injection molding process. This thesis also describes a collaborative effort to demonstrate the feasibility of rapidly produced disposable biochemical diagnostics by using continuous liquid interface production (CLIP) based additive manufacturing (AM). Some unique capabilities of CLIP based AM include the fabrication of 3D devices that can enable compact devices and enhanced mixing and reagent loading compared to traditional 2D microfluidic platforms. Clip based AM can become both the prototyping development material while seamlessly translating to production grade manufacturing at scale. A LAMP based nucleic acid amplification test for the detection of E. coli directly from whole blood is presented with both sample processing, amplification, and detection occurring in forwardly designed CLIP based AM diagnostic cartridge’s in a portable diagnostic cradle. Finally, the thesis also presents the author’s contribution in the development of a new RT-LAMP assay that produces single molecule sensitivity. Patient VTM and saliva samples were used to validate the assay and to confirm its clinical utility. Follow on work will focus on the translation of these RT-LAMP assays in the microfluidic devices and cartridges described earlier.
Issue Date:2021-04-07
Type:Thesis
URI:http://hdl.handle.net/2142/110788
Rights Information:Copyright 2021 Jacob Berger
Date Available in IDEALS:2021-09-17
Date Deposited:2021-05


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