DNA from buccal swab is suitable for rapid genotyping of angiotensin-converting enzyme insertion/deletion (I/D) polymorphism

Angelo Minucci, Giulia Canu, Paola Concolino, Donatella Guarino, Stefania Boccia, Silvana Ficarra, Cecilia Zuppi, Bruno Giardina, Ettore Capoluongo

Research output: Contribution to journalArticlepeer-review


Background: Plasma angiotensin-converting enzyme (ACE) variability between individuals is the results of an insertion/deletion (I. /D) polymorphism in intron 16 of the ACE gene. The I and D alleles differ for the presence or absence of a 288. bp Alu sequence DNA fragment. Methods: The present paper regards the development of a single-tube High Resolution Melting Analysis (HRMA), applied to DNA extracted by buccal swabs, for determining three ACE I/I, I/D, D/D genotypes, in order to obtain a rapid and high throughput method. This method takes advantage of the presence of the 288. bp DNA fragment. Primer design was performed taking into account the possible different efficiency of allele I amplification compared to allele D, avoiding the misclassification of I/D with D/D genotypes. Results: 50 samples previously genotyped by "conventional" PCR protocol already published in literature were 100% concordant with the HRMA results, showing high reproducibility, sensitivity and specificity. ACE genotypes were distinguished by normalized temperature melting curves and by derivate fluorescence plots. Conclusions: HRMA was confirmed as particularly suitable for the identification of ACE I/D polymorphism. Simple setup and rapidity of the analysis (about 1.5. h for 96 samples, including data interpretation) are other important advantages along with low-costs, making this technique useful in clinical research and diagnostics.

Original languageEnglish
Pages (from-to)125-130
Number of pages6
JournalClinica Chimica Acta
Publication statusPublished - Apr 20 2014


  • Angiotensin converting enzyme gene
  • Buccal genomic DNA
  • High resolution melting analysis
  • I/D polymorphism

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Biochemistry, medical
  • Medicine(all)


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