A fast and reliable method for detecting SNP rs67384697 (Hsa-miR-148a binding site) by a single run of allele-specific real-time PCR.

Surface expression of HLA-class I molecules is critical for modulating T/NK lymphocytes' effector functions. Amongst HLA molecules, HLA-C, the most recently evolved form of class I antigens, is subjected to both transcriptional and multiple post transcriptional regulation mechanisms affecting its cell surface expression. Amongst the latter a region placed in the 3' untranslated region (UTR) of HLA-C transcript contains the single nucleotide polymorphism (SNP) rs67384697 "G-ins/del" that has been found to be strictly associated with surface levels of HLA-C allomorphs due to the effect on the binding site of a microRNA (Hsa-miR-148a). Higher expression of HLA-C has proved to influence HIV-1 infection via a better control of viremia and a slower disease progression. More importantly, the analysis of SNP rs67384697 "G-ins/del" combined with the evaluation of the HLA-Bw4/-Bw6 C1/C2 supratype, as well as the KIR genetic asset, has proved to be pivotal in defining the status of Elite Controllers in the caucasian population. Here we describe a new reliable and fast method of allele-specific real-time PCR to monitor the integrity/disruption of the binding site of the microRNA Hsa-miR-148a in a high-throughput format that can be easily applied to studies involving large cohorts of individuals. This article is protected by copyright. All rights reserved.