Comparison of three methods for genotyping of prothrombotic polymorphisms

Marika Bianchi, Enzo Emanuele, Annalisa Davin, Stella Gagliardi, Emanuela Cova, Valentina Meli, Rosita Trotti, Cristina Cereda

Research output: Contribution to journalArticlepeer-review


Several methods have been developed to detect common prothrombotic mutations, including factor V Leiden (G1691), prothrombin G20210A, and methylenetetrahydrofolate reductase (MTHFR) C677C. In this study, we compared the accuracy of three different molecular techniques, i.e.: (1) restriction enzyme digestion (RFLP), (2) real time with hybridization probes and final melting curve (Fluorescence Resonance Energy Transfer, FRET), and (3) real time with hydrolysis probes (TaqMan®). Sequencing was used as the reference standard. Our data showed that RFLPs analysis for the detection of prothrombotic mutations, albeit easy-to-perform, had a limited reliability for assessing correct genotypes. FRET analysis displayed higher resolution than RFLPs. Additionally, FRET analysis was faster and less tedious than sequencing.

Original languageEnglish
Pages (from-to)269-272
Number of pages4
JournalClinical and Experimental Medicine
Issue number4
Publication statusPublished - Dec 2010


  • FRET probes
  • Hydrolysis probes
  • Prothrombotic mutations
  • Real-time PCR
  • RFLPs

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)

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