Developing of primers for amplification of pathogenic variants associated with breast cancer

Abstract

Between 5-10% of cases of breast cancer (BC) are attributed to hereditary genetic mutations. The BCRA 1 and BCRA 2 genes present variants associated with BC with high prevalence and penetrance. The prevalence of these variants differs according to the populations studied, depending on factors such as ethnicity and geographic distribution. Objective: To develop assays for the amplification of regions that have been reported as carriers of genetic variants associated with breast cancer in regional studies. Target regions were selected, primers were designed using the Primer3 tool, PCR amplification of the regions was performed using genomic DNA extracted from a randomly chosen volunteer patient’s whole blood and the amplified fragments were checked on agarose gels. Two primers were designed to amplify regions containing three variants observed in Paraguayan descent women and other countries that may be represented in the Itapuense population: c.5351_5352insA; c.5681_5682insA and c.4035delA. The results of the PCR amplification using the designed primers gave fragments of the expected sizes. The primers designed in this study will allow the evaluation of three variants found in women of Paraguayan descent in Latin American studies.