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Requirements for accurate biosensor detection of anti-lipid biomarker antibodies in active TB
Dissertation (MSc (Biochemistry))--University of Pretoria, 2014.
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| Format: | Thesis |
| Language: | English |
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2026
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| _version_ | 1867613551490760704 |
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| access_status_str | Open Access |
| author2 | Verschoor, J.A. (Jan Adrianus), 1953- |
| author_browse | Verschoor, J.A. (Jan Adrianus), 1953- |
| author_facet | Verschoor, J.A. (Jan Adrianus), 1953- |
| collection | Thesis |
| description | Dissertation (MSc (Biochemistry))--University of Pretoria, 2014. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/110069 |
| institution | University of Pretoria (South Africa) |
| language | English |
| last_indexed | 2026-06-10T12:37:56.779Z |
| license_str | Not specified — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository |
| publishDate | 2026 |
| publishDateRange | 2026 |
| publishDateSort | 2026 |
| record_format | dspace |
| source_str | UPSpace — University of Pretoria Institutional Repository |
| spelling | oai:repository.up.ac.za:2263/110069 Requirements for accurate biosensor detection of anti-lipid biomarker antibodies in active TB Verschoor, J.A. (Jan Adrianus), 1953- mecuino@yahoo.com Ejoh, Vincent Emeka Mycolic acid Tubercolosis Immunoassay Sterol modified lipid Biosensor Dissertation (MSc (Biochemistry))--University of Pretoria, 2014. Much research aiming at eradicating tuberculosis disease is done in the fields of developing management strategies, vaccines, therapeutics and diagnostics. Due to the persistent nature of the causative organism, Mycobacterium tuberculosis, attempts to provide an early, accurate, fast and affordable diagnosis of active disease remains elusive. In our research group a diagnostic test called MARTI (Mycolic acids Antibody Real-time Inhibition) assay is being developed. It uses cell wall mycolic acid from M. tuberculosis as an antigen to detect biomarker antibodies whose prevalence is not hindered by concomitant HIV infection of patients. This project aimed to determine the antigen and antibody requirements needed for the MARTI test. Stereo-controlled chemically synthetic mycolic acids were applied to determine the ideal mycolic acid class composition for antibody detection and find a site on the molecule that can be modified for covalent antigen immobilization without negatively affecting antigenicity. Biomarker antibody sensitivity towards freezing and thawing was assessed as well as the time for antibody exposure to antigen in solution to obtain the best MARTI signal. The results obtained from this study showed that the MARTI-test is best performed on fresh patient serum samples, although a single round of freezing and thawing of serum can be tolerated. Time of exposure of patient serum to MA liposomes during pre-incubation before biosensor analysis also influences the test outcome and required standardization to 20 minutes. The composition of the three MA classes in the MA antigen used for coating the biosensor disc was found to be critically important, with synthetic methoxy-MA proving to be more antigenic than synthetic keto-MA when combined with synthetic _-MA. The synthetic _- and methoxy-MA mixture provided the same MARTI TB diagnostic sensitivity as the natural isolated mixture, but synthetic keto-MA combined with synthetic _-MA sometimes provided false negative MARTI test outcomes. Thiolation of synthetic methoxy-MA on the mycolic motif alpha chain made it less antigenic, but could still be usable as a means whereby this antigen may be covalently linked to the gold biosensor disc in future investigations to improve the reliability of the MARTI-test. Overall, this work provided contributions towards the standardization of the MARTI-test to be considered during its eventual validation for commercialization. This work provides contributions towards the standardization of the MARTI-test to be applied during validation; a strict adherence to fresh serum sample preferably avoiding the need for freezing, a possibility to eventually use synthetic MA covalently bonded to a gold sensor surface for improved consistency of MARTI test results and MA antigen composition in which Methoxy-MA should be more than Keto-MA in the MA mixture. Biochemistry MSc (Biochemistry) 2026-05-15T17:26:13Z 2026-05-15T17:26:13Z 15/01/29 2014 Dissertation http://hdl.handle.net/2263/110069 en application/pdf |
| spellingShingle | Mycolic acid Tubercolosis Immunoassay Sterol modified lipid Biosensor Requirements for accurate biosensor detection of anti-lipid biomarker antibodies in active TB |
| title | Requirements for accurate biosensor detection of anti-lipid biomarker antibodies in active TB |
| title_full | Requirements for accurate biosensor detection of anti-lipid biomarker antibodies in active TB |
| title_fullStr | Requirements for accurate biosensor detection of anti-lipid biomarker antibodies in active TB |
| title_full_unstemmed | Requirements for accurate biosensor detection of anti-lipid biomarker antibodies in active TB |
| title_short | Requirements for accurate biosensor detection of anti-lipid biomarker antibodies in active TB |
| title_sort | requirements for accurate biosensor detection of anti lipid biomarker antibodies in active tb |
| topic | Mycolic acid Tubercolosis Immunoassay Sterol modified lipid Biosensor |
| url | http://hdl.handle.net/2263/110069 |