title: A duplex polymerase chain reaction-restriction fragment length polymorphism for rapid screening of methylenetetrahydrofolate reductase gene variants: Genotyping in acute leukemia.
abstract: BACKGROUND
  Methylenetetrahydrofolate reductase (MTHFR; NM_005957.4) is the key enzyme for folate metabolism which plays in DNA biosynthesis and the epigenetic process of DNA methylation. MTHFR gene polymorphisms, the c. 677C>T and c. 1298A>C have been implicated as risk factors for several types of cancers as the acute leukemia.
  
  
  OBJECTIVE
  We have optimized a duplex polymerase chain reaction-restriction fragment length polymorphism assay (PCR-RFLP) for the simultaneous detection of both variants in acute leukemia patients, from Tunisia.
  
  
  METHODS
  Genomic DNA was extracted from EDTA-anticoagulant blood samples from a total of 50 patients suffering from acute leukemia (AL). After DNA extraction, the polymerase chain reaction using specific primers, designed using Primer 3 Software. Restriction Fragment Length Polymorphism (RFLP) was performed in two separate tubes followed by agarose gel electrophoresis.
  
  
  CONCLUSIONS
  This new method has proved to be a rapid, simple, and reliable method that should facilitate high throughput genotyping of MTHFR polymorphisms in acute leukemia.
authors.affiliation: Laboratory of Histology, Faculty of Medicine of Sfax, University of Sfax, Sfax, Tunisia.
authors.forename: Rim
authors.identifier: http://orcid.org/0000-0003-4415-8785
authors.initials: R
authors.lastname: Frikha
authors.affiliation: Laboratory of Histology, Faculty of Medicine of Sfax, University of Sfax, Sfax, Tunisia.
authors.forename: Nouha
authors.initials: N
authors.lastname: Bouayed
authors.affiliation: Laboratory of Histology, Faculty of Medicine of Sfax, University of Sfax, Sfax, Tunisia.
authors.forename: Bochra
authors.initials: B
authors.lastname: Ben Rhouma
authors.affiliation: Laboratory of Histology, Faculty of Medicine of Sfax, University of Sfax, Sfax, Tunisia.
authors.forename: Leila
authors.initials: L
authors.lastname: Keskes
authors.affiliation: Laboratory of Histology, Faculty of Medicine of Sfax, University of Sfax, Sfax, Tunisia.
authors.forename: Tarek
authors.initials: T
authors.lastname: Rebai
mesh_terms: D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D007938:Leukemia / Q000235:genetics*; D042965:Methylenetetrahydrofolate Reductase (NADPH2) / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length / Q000235:genetics*
chemical_list: D042965:Methylenetetrahydrofolate Reductase (NADPH2)
keywords: methylenetetrahydrofolate reductase; polymerase chain reaction; polymorphisms; restriction fragment length polymorphism