Tuberculosis infects one out of three individuals worldwide. In order to control tuberculosis infection, accurate diagnostics are needed at the point-of-care. An ideal point-of-care diagnostic for tuberculosis would include an integrated system for lysis of the bacteria, extraction of the DNA from the bacterial lysate and the sputum, and detection of a specific biomarker. We compared the use of two different isothermal amplification methods, cross priming amplification (CPA) and loop mediated amplification (LAMP), for the detection of tuberculosis within a previously developed extraction cassette designed for low resource areas. Under ideal laboratory conditions, CPA and LAMP had a limit of detection of 500 copies and 50 copies respectively. As part of an integrated system, CPA and LAMP detected a concentration of bacteria at 1X103 cells/mL at 46 ± 5.8 minutes and 57 ± 4.6 minutes. For the integrated system of tuberculosis detection, CPA generates faster results. However, LAMP was shown to have a lower limit of detection and more specificity under ideal conditions. Overall, this study supports the continued investigation of using isothermal amplification methods combined with a low resource extraction cassette as a point-of-care diagnostic test.
| Identifer | oai:union.ndltd.org:VANDERBILT/oai:VANDERBILTETD:etd-12092013-153815 |
| Date | 09 December 2013 |
| Creators | Creecy, Amy Elizabeth |
| Contributors | David W. Wright, Frederick R. Haselton |
| Publisher | VANDERBILT |
| Source Sets | Vanderbilt University Theses |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.vanderbilt.edu/available/etd-12092013-153815/ |
| Rights | restricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to Vanderbilt University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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