Date of Graduation

7-2021

Document Type

Thesis

Degree Name

Master of Science in Chemical Engineering (MSChE)

Degree Level

Graduate

Department

Chemical Engineering

Advisor/Mentor

Servoss, Shannon L.

Committee Member

Greenlee, Lauren F.

Second Committee Member

Sakon, Joshua

Keywords

Diagnostics; Infectious diseases diagnosis; Loop-mediated isothermal amplification (LAMP); Nucleic Acid Amplification Tests (NAAT); Point-of-Care; Recombinase Polymerase Amplification (RPA)

Abstract

Accurate and early diagnosis of infectious diseases extremely important. Rapid diagnosis allows for effective treatment and increases the chance for recovery without complications. Additionally, the ability to test the populace frequently, swiftly, and affordably significantly aids in containing wide-scale outbreaks. In terms of specificity and sensitivity, nucleic acid amplification tests (NAAT) are one of the best options for diagnosing infectious diseases. Isothermal NAATS present a unique opportunity to create diagnostic tests deployed at a Point-of-Care (POC) level. Specifically, loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) have the potential to deliver reliable POC diagnostics in low-resource settings. When designing POC devices for these NAATs, creating isothermal temperature conditions is perhaps the most significant challenge. This thesis proposes a device capable of incubating either LAMP or RPA in conditions conducive to POC testing. The device can either operate at 58-65°C for ≥60 minutes or 35-42°C for ≥30 minutes; these conditions are required for LAMP or RPA, respectively. This flexibility comes from two factors. Firstly, the energy is provided by a battery-powered polyimide thin-film resistive heater. The voltage conditions can be adjusted depending on the application, but the device does not rely on a microcontroller, which can add intricacy and expense to a device. Secondly, a phase change material (PCM) is used to maintain isothermal conditions via latent energy storage. Usually, chemical heating is regulated via a PCM, and resistive heating is regulated via a microcontroller. This thesis aimed to combine elements of these two common methods to be advantageous for POC testing. To create the device, different heating elements, voltages, PCM amounts, and power sources were tested and optimized for LAMP and RPA. Once the optimal conditions were found, the device was successfully used to perform RPA to amplify DNA.

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