Amplificação gênica alelo-específica na caracterização das hemoglobinas S, C e D e as interaçõ es entre elas e talassemias beta

Abstract

Background: The hemoglobinopathies are a group of hereditary hemoglobin disorders in worldwide distribution, affecting Brazilian population significantly; they are decurrent of alterations in structural genes, responsible for hemoglobin variants, and/or in regulatory genes, resulting the thalassemia. These disorders have been identified in most cases by electrophoretics procedures, and our laboratory experience points out that sometimes they do not obtain enough resolution for a right characterization of mutation. Material and method: The objective of this study was to establish a valid laboratory methodology for the characterization of hemoglobins S, C and D in homozygous or heterozygous and possible interactions, based on the allele-specific genic amplification (PCR-AE) with the use in parallel two primers that differ at their 3′ extremities and are complementary to the normal or mutated sequences. Results and discussion: The results make evident the validation of this methodology in the characterization of these mutations, once this procedure is easy to execute,to reproduce, as well as it is possible to be applied to a significative number of samples.