Investigation into Mycobacterium bovis phenolic glycolipid as a potential biomarker of bovine tuberculosis in urine
2019 Spring. Includes bibliographical references. Mycobacterium 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 t...
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ftmountainschol:oai:mountainscholar.org:10217/195274 2023-06-11T04:06:56+02:00 Investigation into Mycobacterium bovis phenolic glycolipid as a potential biomarker of bovine tuberculosis in urine Morphet, Stephanie Marie Little Thunder Belisle, John Crick, Dean Wilusz, Carol Henry, Chuck 2019-06-14T17:05:34Z born digital doctoral dissertations application/pdf https://hdl.handle.net/10217/195274 English eng eng Colorado State University. Libraries 2000-2019 - CSU Theses and Dissertations MorphetTepp_colostate_0053A_15309.pdf https://hdl.handle.net/10217/195274 Copyright 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. bovine tuberculosis biomarker phenolic glycolipid diagnostic Mycobacterium bovis Text 2019 ftmountainschol 2023-05-06T17:47:17Z 2019 Spring. Includes bibliographical references. Mycobacterium 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 ... Text Antarc* Antarctica Mountain Scholar (Digital Collections of Colorado and Wyoming) |
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bovine tuberculosis biomarker phenolic glycolipid diagnostic Mycobacterium bovis |
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bovine tuberculosis biomarker phenolic glycolipid diagnostic Mycobacterium bovis Morphet, Stephanie Marie Little Thunder Investigation into Mycobacterium bovis phenolic glycolipid as a potential biomarker of bovine tuberculosis in urine |
topic_facet |
bovine tuberculosis biomarker phenolic glycolipid diagnostic Mycobacterium bovis |
description |
2019 Spring. Includes bibliographical references. Mycobacterium 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 ... |
author2 |
Belisle, John Crick, Dean Wilusz, Carol Henry, Chuck |
format |
Text |
author |
Morphet, Stephanie Marie Little Thunder |
author_facet |
Morphet, Stephanie Marie Little Thunder |
author_sort |
Morphet, Stephanie Marie Little Thunder |
title |
Investigation into Mycobacterium bovis phenolic glycolipid as a potential biomarker of bovine tuberculosis in urine |
title_short |
Investigation into Mycobacterium bovis phenolic glycolipid as a potential biomarker of bovine tuberculosis in urine |
title_full |
Investigation into Mycobacterium bovis phenolic glycolipid as a potential biomarker of bovine tuberculosis in urine |
title_fullStr |
Investigation into Mycobacterium bovis phenolic glycolipid as a potential biomarker of bovine tuberculosis in urine |
title_full_unstemmed |
Investigation into Mycobacterium bovis phenolic glycolipid as a potential biomarker of bovine tuberculosis in urine |
title_sort |
investigation into mycobacterium bovis phenolic glycolipid as a potential biomarker of bovine tuberculosis in urine |
publisher |
Colorado State University. Libraries |
publishDate |
2019 |
url |
https://hdl.handle.net/10217/195274 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
2000-2019 - CSU Theses and Dissertations MorphetTepp_colostate_0053A_15309.pdf https://hdl.handle.net/10217/195274 |
op_rights |
Copyright 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. |
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1768379193052102656 |