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Investigation into Mycobacterium bovis phenolic glycolipid as a potential biomarker of bovine tuberculosis in urine

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dc.contributor.authorMorphet, Stephanie Marie Little Thunder, author
dc.contributor.authorBelisle, John, advisor
dc.contributor.authorCrick, Dean, committee member
dc.contributor.authorWilusz, Carol, committee member
dc.contributor.authorHenry, Chuck, committee member
dc.date.accessioned2019-06-14T17:05:34Z
dc.date.available2020-06-10T17:06:51Z
dc.date.issued2019
dc.description.abstractMycobacterium bovis, the etiological agent of bovine tuberculosis (bTB) is reported to cause disease in man and animal alike on every continent aside from Antarctica. Although coordinated efforts have been made for over a century in the US to cease transmission of this pathogen, outbreaks still occur. It has been posited that the failure to eliminate transmission of this pathogen is partially due to the diagnostic in use, which lacks critical sensitivity and specificity. To address this gap, we investigated a potential new method of identifying infected animals that is through the detection of a pathogen-derived biomarker. M. bovis phenolic glycolipid (PGL) is a species-specific, highly abundant, and unique glycolipid that comprises up to 2.5% of the dry cell mass. Coupling an LC-ESI-TOF-MS, method with a solid phase extraction, we successfully detected PGL derived from the urine of naturally-infected cattle. With this knowledge, we aimed to generate a detector of PGL that could be applied in a rapid and field friendly diagnostic platform. Using phage display technology, we selected M13 bacteriophage capable of binding M. bovis PGL with specificity that differentiated between M. bovis PGL and M. Canetti PGL, as well as between total lipid fractions of various species of Mycobacterium and other lipids with similar biochemical properties to PGL. These M. bovis PGL specific phage were able to differentiate between unspiked cattle urine and urine spiked with PGL. Lastly, we assessed the relative stability of PGL, specifically contrasted to another highly abundant mycobacterial glycolipid, phosphatidylinositol mannoside (PIM). We found that PGL was the more stable molecule when testing thermal and chemical stability, as well as when treated with protease K. However, when treating these glycolipids with whole cell lysates derived from fresh bovine organs, accurate detection on an LC-ESI-TOF-MS platform was lost. Further studies will be required to probe the stability of these molecules in vivo. Overall, this potential methodology to assess for infection status may be beneficial in improving the control of M. bovis if further developed, as this new approach has the potential to be more specific and sensitive than the currently used diagnostic.
dc.format.mediumborn digital
dc.format.mediumdoctoral dissertations
dc.identifierMorphetTepp_colostate_0053A_15309.pdf
dc.identifier.urihttps://hdl.handle.net/10217/195274
dc.languageEnglish
dc.language.isoeng
dc.publisherColorado State University. Libraries
dc.relation.ispartof2000-2019
dc.rightsCopyright and other restrictions may apply. User is responsible for compliance with all applicable laws. For information about copyright law, please see https://libguides.colostate.edu/copyright.
dc.subjectbovine tuberculosis
dc.subjectbiomarker
dc.subjectphenolic glycolipid
dc.subjectdiagnostic
dc.subjectMycobacterium bovis
dc.titleInvestigation into Mycobacterium bovis phenolic glycolipid as a potential biomarker of bovine tuberculosis in urine
dc.typeText
dcterms.embargo.expires2020-06-10
dcterms.embargo.terms2020-06-10
dcterms.rights.dplaThis Item is protected by copyright and/or related rights (https://rightsstatements.org/vocab/InC/1.0/). You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).
thesis.degree.disciplineCell and Molecular Biology
thesis.degree.grantorColorado State University
thesis.degree.levelDoctoral
thesis.degree.nameDoctor of Philosophy (Ph.D.)

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