title: [Allele polymorphism analysis in coagulation factors F2, F5 and folate metabolism gene MTHFR by using microchip-based multiplex real time PCR].
abstract: Single nucleotide polymorphism (SNP) genotyping methods are widely used for the detection of hereditary thrombophilias caused by genetic defects in the coagulation system. The hereditary thrombophilias are frequently associated with higher incidences of point mutations in hemostasis (F2 20210G>A, F5 1691G>A) and folate metabolism (MTHFR 677C>Т, MTHFR 1298A>C) genes. Moreover, the combination of gene abnormalities in F2 or/and MTHFR with F5 Leiden mutation leads to increased risk of developing thrombosis. Thus, simultaneous detection of the multiple gene mutations in a sample has important clinical relevance. The microchip-based multiplex real time PCR for estimation of allele specific polymorphism in hemostatic and folate metabolism genes presented here has a high efficiency and may be used for laboratory diagnosis. The optimized protocol for estimation of 4 different types of genetic polymorphisms allowed PCR to be performed with minimal quantity of DNA template and PCR reagents including Taq polymerase and a short-term thermocycling.
authors.affiliation: Institute of Hematology, V.A. Almazov Federal Medical Research Centre, St. Petersburg, Russia; Lumex Group, St. Petersburg, Russia.
authors.forename: K V
authors.initials: KV
authors.lastname: Bogdanov
authors.affiliation: St. Petersburg State University, St. Petersburg, Russia; Lumex Group, St. Petersburg, Russia.
authors.forename: M M
authors.initials: MM
authors.lastname: Nikitin
authors.affiliation: St. Petersburg State University, St. Petersburg, Russia; Lumex Group, St. Petersburg, Russia.
authors.forename: M N
authors.initials: MN
authors.lastname: Slyadnev
mesh_terms: D005165:Factor V / Q000235:genetics*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D042965:Methylenetetrahydrofolate Reductase (NADPH2) / Q000235:genetics*; D060885:Multiplex Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D011516:Prothrombin / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D019851:Thrombophilia / Q000235:genetics*
chemical_list: D005165:Factor V; D011516:Prothrombin; C573423:MTHFR protein, human; D042965:Methylenetetrahydrofolate Reductase (NADPH2)
keywords: F2; F5; MTHFR; PCR; hereditary thrombophilias; single nucleotide polymorphism