Real time PCR assay with fluorescent hybridization probes for genotyping intronic polymorphism in presenilin-1 gene

Clin Chim Acta. 2006 Feb;364(1-2):343-4. doi: 10.1016/j.cccn.2005.06.017. Epub 2005 Sep 1.

Abstract

Background: Identification of all susceptibility loci for Alzheimer's disease has been a major goal in resolving the pathogenesis of this disease.

Methods: A PCR assay with fluorescently labeled oligonucleotide hybrinization probes with subsequent fluorescent probe melting point analysis was developed.

Results: Allelic discrimination of intronic polymorphism of presenilin-1 gene and the restriction fragment length polymorphism method yielded identical results, proving its usefulness for genotyping PS1 gene.

Conclusions: This method provides excellent robustness, speed, and accuracy, and is well suited for determination of the polymorphism in both small and large numbers of samples. This assay could help to overcome the controversy regarding the association between the PS1 s165932 intronic polymorphism and Alzheimer's disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Alzheimer Disease / genetics
  • DNA / chemistry
  • DNA / genetics
  • DNA Mutational Analysis / methods
  • DNA Probes / chemistry
  • DNA Probes / genetics*
  • Fluorescent Dyes / chemistry
  • Genetic Predisposition to Disease / genetics
  • Genotype
  • Humans
  • Introns / genetics*
  • Membrane Proteins / genetics*
  • Nucleic Acid Denaturation
  • Nucleic Acid Hybridization / methods
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Restriction Fragment Length
  • Polymorphism, Single Nucleotide*
  • Presenilin-1
  • Transition Temperature

Substances

  • DNA Probes
  • Fluorescent Dyes
  • Membrane Proteins
  • PSEN1 protein, human
  • Presenilin-1
  • DNA