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

BACKGROUND: 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. METHODS: 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. RESULTS: 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%. CONCLUSIONS: Prevalidation procedure described in this paper can be applied, not only to the allele quantification, but also to many other real time PCR applications.