title: Evolving methods for single nucleotide polymorphism detection: Factor V Leiden mutation detection.
abstract: BACKGROUND
  The many techniques used to diagnose the Factor V Leiden (FVL) mutation, the most common hereditary hypercoagulation disorder in Eurasians, and the most frequently requested genetic test reflect the evolving strategies in protein and DNA diagnosis.
  
  
  METHODS
  Here, molecular methods to diagnose the FVL mutation are discussed.
  
  
  RESULTS
  Protein-based detection assays include the conventional functional activated protein C resistance coagulation test and the recently reported antibody-mediated sensor detection; and DNA-based assays include approaches that use electrophoretic fractionation e.g., restriction fragment length polymorphism, denaturing gradient gel electrophoresis, and single-stranded conformational PCR analysis, DNA hybridization (e.g., microarrays), DNA polymerase-based assays, e.g., extension reactions, fluorescence polarization template-directed dye-terminator incorporation, PCR assays (e.g., amplification-refractory mutation system, melting curve analysis using real-time quantitative PCR, and helicase-dependent amplification), DNA sequencing (e.g., direct sequencing, pyrosequencing), cleavase-based Invader assay and ligase-based assays (e.g., oligonucleotide ligation assay and ligase-mediated rolling circle amplification).
  
  
  CONCLUSIONS
  The method chosen by a laboratory to diagnose FVL not only depends on the available technical expertise and equipment, but also the type, variety, and extent of other genetic disorders being diagnosed.
authors.affiliation: Molecular Biotechnology Research Laboratory, Boston University, Boston, MA, USA. hoh@bu.edu
authors.forename: Herin
authors.initials: H
authors.lastname: Oh
authors.forename: Cassandra L
authors.initials: CL
authors.lastname: Smith
mesh_terms: D001780:Blood Coagulation Tests; D004252:DNA Mutational Analysis / Q000379:methods*; D005165:Factor V / Q000235:genetics*; D006801:Humans; D007118:Immunoassay; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D012150:Polymorphism, Restriction Fragment Length
chemical_list: C095381:factor V Leiden; D005165:Factor V