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. © 2010 Springer-Verlag.
CITATION STYLE
Bianchi, M., Emanuele, E., Davin, A., Gagliardi, S., Cova, E., Meli, V., … Cereda, C. (2010). Comparison of three methods for genotyping of prothrombotic polymorphisms. Clinical and Experimental Medicine, 10(4), 269–272. https://doi.org/10.1007/s10238-010-0096-3
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