Limitations of commercial RNA extraction kits make them inappropriate for use in a clinical setting. This project develops and evaluates a point-of-care process for simple and rapid extraction of RNA from clinical samples. The novel RNA extraction method does not require laboratory equipment and is simpler and faster. In this thesis the proposed method is compared to three existing commercial extraction kits. Comparisons were made using RT-PCR of extraction solutions obtained after adding a known number of copies of respiratory syncytial virus (RSV) RNA to buffer or negative patient samples and using previously stored RSV positive patient samples. A comparison of viral RNA extraction from buffer found that our initial design of the proposed method achieved extraction efficiency of 80 to 90% of commercial kits. Extraction performance using previously stored RSV positive patient samples ranged from ~15 to 67% of commercial kits. With further optimization this approach would likely have a significant impact on moving genome-based detection methods into a clinical setting.
| Identifer | oai:union.ndltd.org:VANDERBILT/oai:VANDERBILTETD:etd-09022009-110506 |
| Date | 02 September 2009 |
| Creators | Klemm, Amy Song |
| Contributors | Professor F.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-09022009-110506/ |
| Rights | unrestricted, 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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