Microfludic point of care platforms for the detection of viruses, proteins, and nucleic acids

Jankelow, Aaron

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https://hdl.handle.net/2142/124195 Copy

Description

Title

Microfludic point of care platforms for the detection of viruses, proteins, and nucleic acids

Author(s)

Jankelow, Aaron

Issue Date

2024-04-15

Director of Research (if dissertation) or Advisor (if thesis)

• Bashir, Rashid
• Valera, Enrique

Doctoral Committee Chair(s)

• Bashir, Rashid
• Valera, Enrique

Committee Member(s)

• Cunningham, Brian
• Smith, Andrew

Department of Study

Bioengineering

Discipline

Bioengineering

Degree Granting Institution

University of Illinois at Urbana-Champaign

Degree Name

Ph.D.

Degree Level

Dissertation

Keyword(s)

• Microfluidics
• Point of Care
• LAMP
• Coulter Counting
• Pathogen Detection
• Virus detection
• Diagnostics
• Immunoassays
• hydrogel beads

Abstract

Point-of-care diagnostics are critical to the advancement of patient healthcare by allowing for rapid and sensitive diagnoses that will allow patients to get better care as fast as possible, particularly in low resource areas where access to hospital labs may be poor or during epidemic and pandemic situations where rapid diagnostics that can be done with as little travel as possible will be vital to managing the outbreak and mitigating the damages caused by the diseases. Microfluidics platforms have proven valuable in the advancement of point-of-care diagnostics by allowing for portable devices that can perform entire assays with minimal training from the user. This thesis describes a point-of-care platform for the diagnosis of Zika Virus from whole blood using RT-LAMP that can be performed on a portable platform and read with a smartphone camera. An RT-LAMP assay was first designed and validated off-chip in buffer with Zika genomic RNA and gamma-irradiated samples of zika virus. This assay was then validated off-chip in whole blood, before we tested the on-chip mixing of our microfluidic mixing module and validated its performance compared to off-chip mixing. Finally, we performed the entire assay on-chip with mixing done on our mixing module and amplification performed on our amplification module which was inserted into our reading and heating platform and recorded on a smartphone camera. The smartphone data was then analyzed using our proprietary app, using both a standard global analysis of the entire amplification chamber as well as our novel Spatial-LAMP approach, in order to determine if and when amplification occurred. Additionally, this thesis also describes advancements made in an existing electrical differential detection platform that used the capture of beads between an entrance and exit count in order to detect the presence of specific biomarkers in a biological solution. Multiplexing capabilities were incorporated onto this existing platform by utilizing different beads with distinct electrical signals in order to allow for more robust and versatile assays to be developed with this platform. First, we demonstrated the ability of this platform to achieve multiplexed capture of two sepsis biomarkers, IL-6 and PCT, from plasma, which we validated against flow cytometry results. Next, we did work incorporating the use of Hydrogel Beads to allow for a greater multiplexing capacity than what could be achieved with commercially available microbeads and demonstrated their uses in multiplexed capture with protein and nucleic acid targets. After this we demonstrated the capability of the system to expand the library of available targets that this platform can detect to include whole viruses by demonstrating the multiplexed capture of Hepatitis A Virus and Human Parvovirus B19 from plasma. Finally, we discuss an ongoing project to further increase the multiplexing capacity of our electrical differential platform to be able to detect up to 10 different biomarkers by utilizing a set of five nondegradable beads that can each be paired with a degradable counterpart, which can then be differentiated through the addition of a degradation chamber that selectively degrades the degradable beads while leaving the nondegradable beads intact, and a post-degradation count that when paired with the counts before and after capture will allow us to determine the capture of all ten beads independently.

Graduation Semester

2024-05

Type of Resource

Thesis

Copyright and License Information

Copyright 2024 Aaron Jankelow

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