Small Amplicons High Resolution Melting Analysis (SA-HRMA) allows successful genotyping of acid phosphatase 1 (ACP1) polymorphisms in the Italian population

BackgroundThe ACP1 gene, encoding a low-molecular-weight phosphotyrosine phosphatase (LMW-PTP), has been suggested as a common genetic factor of several human diseases, including inflammatory and autoimmune diseases, favism and tumors. For this reason, the ACP1 enzyme has been investigated by case–control studies for decades. Initially based on protein electrophoresis, the ACP1 phenotype is now determined by DNA-based techniques.MethodsHere, we report a rapid optimized method which employs HRMA for ACP1 polymorphism identification, a molecular approach that we used to screen 80 healthy Italian subjects.ResultsHRMA proved particularly suitable for detecting ACP1 genotypes. In fact, HRMA results were 100% concordant with direct sequencing. In addition, ACP1 genotype frequency in the Italian population was in accordance with the literature [4% (*A/A), 36% (*A/B), 4% (*A/C), 50% (*B/B), 6% (*B/C)].ConclusionsHRMA was found to be a simple, rapid, sensitive and low cost method potentially useful in research and diagnostic laboratories. Finally, use of small amplicons for the set-up allowed us a better optimization of HRMA. For this reason, we present such an approach as small amplicons high resolution melting analysis (SA-HRMA).Finally, ACP1 genotype frequency in the Italian population reported in this study may contribute to a better interpretation of ACP1 allelic frequency variation.

► ACP1 gene as common genetic factor of several human diseases.
► ACP1 polymorphisms screening based on HRMA is found to be a simple, rapid, sensitive and low cost.
► The use of Small Amplicons allowed to better characterize HRMA approach as SA-HRMA.
► We found that Italian ACP1 genotypes frequency follows the Hardy-Weinberg equilibrium.