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The prevalence of HIV primary resistance mutations down to the level of 1% minor variants was investigated using ultradeep pyrosequencing (UDPS) in HIV-positive Korean blood donors and in treatment naïve chronic patients for the comparison.
This is in concordance with recent MPS studies that detected mutations down to 1% [ 4, 8].
Furthermore, melting curve analysis immediately after PCR can identify small mutations, down to single base changes.
All mutations down to 6.6% allele frequency could be detected with 100% specificity.
In contrast, pyrosequencing detected 100% of the mutations down to 5% allele frequency but exhibited only 90% specificity.
After whittling the mutations down to those that are recurrent, information about therapies and prognostic information can be found at a number of locations.
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Pyrosequencing detected the p.V600E mutation down to 5% mutated alleles in a background of wildtype alleles but with values for the relative fluorescent units close to our threshold of 30.
The researchers tracked the mutation down to a gene called fibroblast growth factor receptor 1 (fgfr1).
Similar results were obtained using the TheraScreen K-RAS Mutation kit (mutation detected down to 1.25% tumor cells) consistent with the reported sensitivity of this method (data not shown).
In patients who had BART as an elective test, LRs made up 5.9% of all mutations, breaking down to 11.8% of BRCA1 and 1.2% of BRCA2 mutations.
Our snapback primer system, with a robust reproducibility, can also discriminate the JAK2 V617F and MPL W515L/K mutations simultaneously down to the 0.1% level.
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