Development of a real-time PCR incorporating high resolution melting analysis to screen HIV-1 samples for resistance-related codons

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dc.contributor.author Sacks, David
dc.date.accessioned 2011-02-01T09:47:16Z
dc.date.available 2011-02-01T09:47:16Z
dc.date.issued 2011-02-01
dc.identifier.uri http://hdl.handle.net/10539/8993
dc.description MSc (Med), Virology, Faculty of Health Sciences, University of the Witwatersrand en_US
dc.description.abstract Introduction High resolution melting analysis (HRMA) accurately, rapidly and cost effectively detects single nucleotide polymorphisms by monitoring DNA dissociation kinetics. This technology was applied to HIV samples to assess whether it could be used to detect clinically relevant drug resistance mutations. Methods HRMA-PCR assays incorporating unlabeled probes were designed to genotype 12 mutation codons in the HIV-1 p66/p51 of engineered plasmids and 116 HIV-1 samples. Results HRMA correctly genotyped 63%-88% of the K103N, Y181C, M184V, Q151M and G190A mutations. Each assay had a 1.7%-3.4% discordance, most of which was due to the increased analytical sensitivity of HRMA (~5-20%). Only mutant K65R and V106M were correctly identified while the 41, 67, 70, 215 and 225 codons could not be genotyped. Assay modifications had some success in masking the affects of polymorphisms. Conclusion These assays can be used for genotyping selected major HIV-1 resistance mutations and should be further developed as a resistance surveillance tool. en_US
dc.language.iso en en_US
dc.subject HIV-1 en_US
dc.subject mutations en_US
dc.subject drug resistance en_US
dc.title Development of a real-time PCR incorporating high resolution melting analysis to screen HIV-1 samples for resistance-related codons en_US
dc.type Thesis en_US


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