title: Evaluation of high-resolution melting analysis for screening the LDL receptor gene.
abstract: OBJECTIVE
  High-resolution melting analysis has been validated for screening several genes of clinical significance. We evaluated whether it would be useful for detecting mutations in the LDL receptor gene (LDLR).
  
  
  METHODS
  We analysed 60 samples heterozygous for different LDLR sequence variants, and compared the melting curves to those of the corresponding homozygous normal amplicons to assess whether they would be detected in a mutation scanning screen using melting analysis.
  
  
  RESULTS
  Although six mutations in exon 4 did not yield usable melting data, 96% (52/54) of the other variants were detected by the melting analysis, while two variants were poorly resolved.
  
  
  CONCLUSIONS
  Melting analysis is a rapid and sensitive mutation scanning technique, but is not suitable for some GC-rich amplicons such as LDLR exon 4, or exons with high frequencies of benign polymorphisms which would require follow-up DNA sequencing. We developed a successful strategy which combines melting analysis with direct sequencing of problematic amplicons.
authors.affiliation: Molecular Pathology, Canterbury Health Laboratories, Christchurch, New Zealand. andrew.laurie@cdhb.govt.nz
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Laurie
authors.forename: Peter M
authors.initials: PM
authors.lastname: George
mesh_terms: D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis*; D005820:Genetic Testing / Q000379:methods*; D014644:Genetic Variation; D006801:Humans; D006938:Hyperlipoproteinemia Type II / Q000175:diagnosis / Q000235:genetics; D009154:Mutation; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D020033:Protein Isoforms / Q000032:analysis / Q000235:genetics; D011973:Receptors, LDL / Q000032:analysis / Q000235:genetics*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D044366:Transition Temperature
chemical_list: D020033:Protein Isoforms; D011973:Receptors, LDL; D004247:DNA