Biomolecular Detection Assays

Simple item page
dc.contributor.advisorWillson, Richard C.
dc.contributor.committeeMemberOrman, Mehmet A.
dc.contributor.committeeMemberKourentzi, Katerina
dc.contributor.committeeMemberShevkoplyas, Sergey S.
dc.contributor.committeeMemberMohan, Chandra
dc.creatorHlavinka, Victoria
dc.date.accessioned2023-01-11T17:06:42Z
dc.date.createdMay 2022
dc.date.issued2022-05-10
dc.date.updated2023-01-11T17:06:43Z
dc.description.abstractIn the present work, three methods for the detection or monitoring of biomolecules are outlined in detail. The first study describes the development of a saliva-based enzymatic screening assay for diabetes mellitus. In the second study, lateral flow assays (LFAs) utilizing commercial colloidal gold and blue latex nanoparticle reporters are compared to persistent luminescence nanoparticle (PLNP; “nanophosphor”) LFAs to assess the sensitivity of nanophosphors as a reporter system. The final study demonstrates the utility of a nanophosphor-based LFA for detecting low concentrations of dengue virus (DENV) biomarker non-structural protein 1 (NS1). 1,5-Anhydroglucitol (AHG) is a naturally occurring monosaccharide and a clinically validated blood biomarker for diabetes. The blood concentration of AHG falls during periods of hyperglycemia, as glucose outcompetes AHG for kidney reuptake. Salivary AHG quantification has been suggested to be useful for diabetes screening but has not been implemented in any widely applicable fashion. We have developed a chemiluminescence assay to quantify AHG in saliva and demonstrated that the assay could distinguish between healthy and diabetic individuals (N = 265; p < 0.0001, ROC AUC = 0.82). These findings suggest that, with further validation, this approach may serve as the basis of a non-invasive tool for diabetes screening. Commercially-available LFAs commonly use colloidal gold or blue latex nanoparticles reporter systems that lack sensitivity and are prone to human error when interpreted visually. We have developed nanophosphors that can detect low levels of antigen. In a comparison study, a nanophosphor-based human immunoglobulin G (IgG) LFA had a limit of detection of 0.625 ng/mL, an 81-fold and 58-fold increase in sensitivity over colloidal gold and blue latex nanoparticles, respectively. Current DENV diagnostic methods are commonly unspecific and cannot detect early infection. We have developed an inexpensive, rapid LFA to detect DENV NS1, a known marker of early dengue infection. Using strontium aluminate nanophosphors as reporters, we achieved a limit of detection of 1 ng/mL DENV serotype 1 NS1 antigen. Our assay is comparable to a laboratory-based NS1 ELISA with a 1 ng/mL limit of detection.
dc.description.departmentChemical and Biomolecular Engineering, William A. Brookshire Department of
dc.format.digitalOriginborn digital
dc.format.mimetypeapplication/pdf
dc.identifier.urihttps://hdl.handle.net/10657/13299
dc.language.isoeng
dc.rightsThe author of this work is the copyright owner. UH Libraries and the Texas Digital Library have their permission to store and provide access to this work. Further transmission, reproduction, or presentation of this work is prohibited except with permission of the author(s).
dc.subjectDiabetes mellitus
dc.subject1,5-anhydroglucitol
dc.subjectLateral flow assays
dc.subjectPersistent luminescence
dc.subjectDengue virus
dc.titleBiomolecular Detection Assays
dc.type.dcmiText
dc.type.genreThesis
dcterms.accessRightsThe full text of this item is not available at this time because the student has placed this item under an embargo for a period of time. The Libraries are not authorized to provide a copy of this work during the embargo period.
local.embargo.lift2024-05-01
local.embargo.terms2024-05-01
thesis.degree.collegeCullen College of Engineering
thesis.degree.departmentChemical and Biomolecular Engineering, William A. Brookshire Department of
thesis.degree.disciplineChemical Engineering
thesis.degree.grantorUniversity of Houston
thesis.degree.levelDoctoral
thesis.degree.nameDoctor of Philosophy

Files

Original bundle

Now showing 1 - 1 of 1
Thumbnail Image
Name:
HLAVINKA-DISSERTATION-2022.pdf
Size:
1.78 MB
Format:
Adobe Portable Document Format
Download

License bundle

Now showing 1 - 2 of 2
No Thumbnail Available
Name:
PROQUEST_LICENSE.txt
Size:
4.43 KB
Format:
Plain Text
Description:
Download
No Thumbnail Available
Name:
LICENSE.txt
Size:
1.82 KB
Format:
Plain Text
Description:
Download

Collections

Published ETD Collection