Whether this reflects a somewhat lower sensitivity of our methodology compared to Juliano et al remains to be determined, but it is certainly consistent with the conclusion that these recrudescences are not associated with selection of a minority, mutant Pfcrt clone.
For this reason, we lowered the T c to 79.4°C to further enhance amplification of minority mutant alleles and confirm the KRAS mutations we detected in the 4 discrepant samples.
The second assumption is that minority mutants have the same weight in genotypic susceptibility interpretation algorithms as if they were present at higher levels.
Therefore, to maximize specificity, we used pre-antiretroviral (pre-ARV) drug era wildtype virus samples to define cutoffs that exclude the detection of naturally-occurring minority mutants.
In Le's study, however, samples were collected before etravirine, darunavir or raltegravir became available, so the effect additional minority mutants on HIV susceptibility to these drugs could not be evaluated.
Additional minority mutants increased a subject's genotypic resistance to one or more antiretrovirals in 17 of 22 individuals (77%), correlated with the failing drugs in 21% subjects, and with historical antiretroviral use in 79% subjects.
Whereas most studies addressing the role of minority variants have been performed in antiretroviral-naïve subjects [2], [3], [4], [5], [6], [7], [9], [10], [11], [12], [13], less information exists on the clinical significance of minority mutants in antiretroviral-experienced individuals.
To detect a minority of mutant KRAS alleles among abundant wild-type alleles, we developed a sensitive DNA sequencing assay using Pyrosequencing, ie, nucleotide extension sequencing with an allele quantification capability.
However, this phenotype was not the same in every cell as a minority of mutant-transfected cells appeared to show some Nav1.7 staining at the plasma membrane.
We have reconstructed a foot-and-mouth disease virus (FMDV) quasispecies, with two matched pairs of distinguishable mAb-escape mutants as minority genomes of the mutant spectrum.
In addition, a minority of the mutant notch3 mRNAs contained aberrantly spliced sequences that result in partial deletions of exon 13.
