The facile detection of the nt 1226 mutation of glucocerebrosidase by 'mismatched' PCR

Clin Chim Acta. 1990 Dec 24;194(2-3):161-6. doi: 10.1016/0009-8981(90)90130-k.


The most common Gaucher disease-producing mutation among Ashkenazi Jews is an A----G substitution at cDNA nt 1226 (genomic nt 5841). We describe a simple method for detecting this mutation both in genomic DNA and in cDNA by performing polymerase chain reaction (PCR) using a 5'-primer mismatched at one nucleotide so as to create an Xho I restriction site. When the mutation is present. the 105 bp fragment formed is cleaved to 89 and 16 nt fragments. The 89 bp fragment is easily visualized on a gel making it possible to distinguish individuals who do not have the mutation from heterozygotes and homozygotes.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • Deoxyribonucleases, Type II Site-Specific
  • Gaucher Disease / enzymology*
  • Glucosylceramidase / genetics*
  • Heterozygote
  • Homozygote
  • Humans
  • Molecular Sequence Data
  • Mutation*
  • Polymerase Chain Reaction*


  • CTCGAG-specific type II deoxyribonucleases
  • Deoxyribonucleases, Type II Site-Specific
  • Glucosylceramidase