Evolving methods for single nucleotide polymorphism detection: Factor V Leiden mutation detection

• Published in: Journal of clinical laboratory analysis 2011, Vol.25 (4), p.259-288
• Main Authors: Oh, Herin, Smith, Cassandra L.
• Format: Article
• Language: English
• Subjects: activated protein C; amplification-refractory mutation system; Blood Coagulation Tests; Coagulation; Coagulation factors; coagulation tests; denaturing gradient gel electrophoresis; DNA assay; DNA microarrays; DNA Mutational Analysis - methods; DNA sequencing; ELISA; Factor V; Factor V - genetics; Factor V Leiden; Fluorescence polarization; fluorescence polarization template-directed dye-terminator incorporation; Gel electrophoresis; gold particle probes; helicase-dependent amplification; Humans; immuno-optical sensor; Immunoassay; invader; ligase-mediated rolling circle amplification; Melting curve; melting curve analysis using real-time quantitative PCR; microarrays; Molecular Diagnostic Techniques - methods; Mutation; Oligonucleotide Array Sequence Analysis; oligonucleotide ligation assay; Oligonucleotides; Original; Polymerase chain reaction; Polymorphism, Restriction Fragment Length; Restriction fragment length polymorphism; sequencing; single-base extension; Single-nucleotide polymorphism; single-strand conformation polymorphism
• ISSN: 0887-8013; 1098-2825; 1098-2825
• DOI: 10.1002/jcla.20470

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). Conclusion: 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. J. Clin. Lab. Anal. 25:259–288, 2011. © 2011 Wiley‐Liss, Inc.