A novel TaqMAMA assay for allelic discrimination of immunoproteasome subunit PSMB8 in pediatric patients

Abstract

A more recent improvement in MAMA was made when the allele selectivity of MAMA primers was combined with the 5' fluorogenic exonuclease (TaqMan) assay. This strategy referred to as TaqMAMA. A SNP rs9357155 at the PSMB8 locus (GenBank access number NM_148919.3.) on chromosome 6 showing a C/T transition in position 32842071, has been chosen as model in this study. Tested assays have provided to be linear over two log10 of magnitude and have efficiencies close to the average of 80.3%. Average interference limit values of TaqMAMA have been considered around the 0.6%. Coefficient of variation (CV) for PSMB8c and t were approximatively 1%. Prevalidation procedure described in this paper can be applied, not only to the allele quantification, but also to many other real time PCR applications.