Published in:
01-12-2010 | Short Communication
Comparison of three methods for genotyping of prothrombotic polymorphisms
Authors:
Marika Bianchi, Enzo Emanuele, Annalisa Davin, Stella Gagliardi, Emanuela Cova, Valentina Meli, Rosita Trotti, Cristina Cereda
Published in:
Clinical and Experimental Medicine
|
Issue 4/2010
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Abstract
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.