Small amplicons high resolution melting analysis (SA-HRMA) allows successful genotyping of acid phosphatase 1 (ACP1) polymorphisms in the Italian population

Clin Chim Acta. 2013 Feb 1:416:86-91. doi: 10.1016/j.cca.2012.11.023. Epub 2012 Nov 29.

Abstract

Background: The 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.

Methods: Here, 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.

Results: HRMA 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)].

Conclusions: HRMA 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.

MeSH terms

  • Acid Phosphatase / genetics*
  • Base Sequence
  • DNA Mutational Analysis / economics
  • DNA Mutational Analysis / standards
  • Genotype
  • Humans
  • Italy
  • Molecular Sequence Data
  • Nucleic Acid Amplification Techniques / economics
  • Nucleic Acid Amplification Techniques / standards*
  • Polymorphism, Genetic*
  • Reproducibility of Results
  • Time Factors

Substances

  • Acid Phosphatase