Thesis advisor: Gregory R. Chiklis / Thesis advisor: Kathleen Dunn / In molecular diagnostics, the polymerase chain reaction (PCR) is used to amplify small amounts of nucleic acids found in patient samples, allowing for detection of diseases within hours of infection. This early detection allows medical professionals to diagnose and treat patients with greater success. It is crucial that internal controls, such as NATtrol™-treated microorganisms, are used in these PCR assays to avoid false-negative results and ensure accurate diagnosis of patients. NATtrol™ treatment renders microorganisms non-infectious while leaving them fully intact with their complete RNA or DNA genomes. Therefore, NATtrol™-treated microorganisms can be used in PCR as full-process internal controls that are spiked into patient samples and co-extracted and co-amplified within the sample. If the spiked NATtrol™ control returns expected results on the test, then the patient sample result can also be trusted. Here, we performed studies to validate the use of NATtrol™-treated MS2 virus as a universal full-process internal molecular control. In these studies, a quantitative, real-time, reverse-transcription PCR (qRT-PCR) assay was performed on the Roche LightCycler 480 instrument. Studies included working range validation, limit of detection, within-run precision, between-run precision, real-time stability, freeze-thaw (transport) stability, and open-vial (use-life) stability. All studies demonstrated the precision and stability of the MS2 NATtrol™ molecular control. / Thesis (BS) — Boston College, 2014. / Submitted to: Boston College. College of Arts and Sciences. / Discipline: College Honors Program. / Discipline: Biology Honors Program. / Discipline: Biology.
| Identifer | oai:union.ndltd.org:BOSTON/oai:dlib.bc.edu:bc-ir_102389 |
| Date | January 2014 |
| Creators | McGlynn, Kayleigh Erin |
| Publisher | Boston College |
| Source Sets | Boston College |
| Language | English |
| Detected Language | English |
| Type | Text, thesis |
| Format | electronic, application/pdf |
| Rights | Copyright is held by the author, with all rights reserved, unless otherwise noted. |
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