Abstract

In recent years, the world has faced numerous pandemics caused by viral diseases, leading to considerable morbidity, mortality, and economic disruptions. Developing vaccines for pandemics can be a challenging task due to the rapid mutation of viruses and the significant investment of time required for their development. While real-time polymerase chain reaction and immunoassay techniques are frequently utilized for viral diagnostics, they possess inherent limitations. Therefore, there is a pressing need to rapidly and accurately identify viral infections to manage diseases effectively. This work explores the use of nanotechnology and clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) proteins to achieve rapid and accurate viral identification. One biosensing assay and two microdevices are presented, each with innovative features. The biosensing assay employs Quantum dots as indicators and Cas-mediated nucleic acid probe cleavage to provide a simple “Yes-or-No” readout. By adding a magnetic bead washing step, the assay delivers comparable detection sensitivity in both buffer and plasma, largely eliminating high background issues raised in other publications. One of the microdevices presented couples the isothermal amplification technique with the CRISPR-Cas assay, achieving ultra-sensitivity with the help of a hand warmer pouch for heating. This microdevice is pre-loaded with reagents and can be operated by simply pulling rods, making it possible for ordinary people to perform on-site diagnostics. The other microdevice incorporates high-aspect-ratio micropillars and the work demonstrates that using high-density micropillars can achieve higher detection sensitivity.

Library of Congress Subject Headings

CRISP activation; DNA viruses--Identification; Quantum dots

Publication Date

5-2023

Document Type

Dissertation

Student Type

Graduate

Degree Name

Mechanical and Industrial Engineering (Ph.D)

Department, Program, or Center

Mechanical Engineering (KGCOE)

Advisor

Ke Du

Advisor/Committee Member

Andre O. Hudson

Advisor/Committee Member

Karin Wuertz-Kozak

Comments

This dissertation has been embargoed. The full-text will be available on or around 2/21/2024.

Campus

RIT – Main Campus

Plan Codes

MIE-PHD

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