title: Primer-engineered multiplex PCR-RFLP for detection of MTHFR C677T, prothrombin G20210A and factor V Leiden mutations.
abstract: Single-nucleotide polymorphisms in the genes that code for coagulation factors V (factor V Leiden) and II (prothrombin, G20210A), as well as the methylenetetrahydrofolate reductase (MTHFR, C677T) gene, have been implicated in the majority of cases of hereditary thrombophilia. We have developed a multiplex PCR-RFLP assay based on MnlI endonuclease digestion for the simultaneous detection of mutations in the FV, FII, and MTHFR genes. Digested amplification products were analyzed by gel electrophoresis in a single gel lane and visualized by ethidium bromide. This approach is a rapid and convenient method, hence economic, that alternate to others described for the detection of FVL, G20210A and C677T mutations.
authors.affiliation: Burc Molecular Diagnostic Laboratory, Istanbul, Turkey.
authors.forename: Vedat
authors.initials: V
authors.lastname: Koksal
authors.forename: Ibrahim
authors.initials: I
authors.lastname: Baris
authors.forename: Ozdal
authors.initials: O
authors.lastname: Etlik
mesh_terms: D000483:Alleles; D003596:Cytosine / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics*; D005165:Factor V / Q000032:analysis* / Q000235:genetics; D006147:Guanine / Q000378:metabolism; D006801:Humans; D042965:Methylenetetrahydrofolate Reductase (NADPH2) / Q000032:analysis* / Q000235:genetics; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011516:Prothrombin / Q000032:analysis* / Q000235:genetics
chemical_list: D017931:DNA Primers; C095381:factor V Leiden; D006147:Guanine; D003596:Cytosine; D005165:Factor V; D011516:Prothrombin; D042965:Methylenetetrahydrofolate Reductase (NADPH2)