R117H and IVS8-5T cystic fibrosis mutation detection by restriction enzyme digestion

Mol Diagn. 2000 Sep;5(3):235-8. doi: 10.1054/modi.2000.9730.

Abstract

The number of thymines in a thymine repeat in intron 8 (IVS8) next to the exon 9 of the cystic fibrosis gene (CFTR) affects the efficiency of this site to act as a splice acceptor site and the subsequent inclusion or skipping of exon 9 into the CFTR protein. Another source of genetic variation, responsible for mild cystic fibrosis (CF) and/or congenital bilateral absence of the vas deferens, is the mutation R117H, located in exon 4 of CFTR. Both these genetic variants can be detected using restriction site-generating PCR amplification. The severity of the CF phenotype is partly dependent on the IVS8 background on which R117H occurs; thus, it is important to be able to test clinically for both these variants. A method using restriction digestion of PCR product is presented.

MeSH terms

  • Cystic Fibrosis / diagnosis*
  • Cystic Fibrosis / genetics*
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics*
  • DNA Restriction Enzymes / chemistry*
  • Genetic Markers
  • Humans
  • Introns
  • Point Mutation*
  • Polymerase Chain Reaction*
  • Polymorphism, Genetic
  • Repetitive Sequences, Nucleic Acid

Substances

  • CFTR protein, human
  • Genetic Markers
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • DNA Restriction Enzymes