Determination of Lewis FUT3 gene mutations by PCR using sequence-specific primers enables efficient genotyping of clinical samples

Hum Mutat. 2001 Oct;18(4):358-9. doi: 10.1002/humu.1204.

Abstract

We have developed a polymerase chain reaction method using sequence-specific primers (PCR-SSP) for rapid and correct genotyping of the common Lewis (FUT3) gene mutations 59T>G, 202T>C, 314C>T, 508G>A, and 1067T>A. The PCR-SSP method was validated on 20 healthy blood donors and 16 non-insulin-dependent diabetic patients. All individuals were in parallel genotyped by our established polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. The FUT3 genotypes, determined with the PCR-SSP method, were in complete accordance with the results of the PCR-RFLP reference method. The PCR-SSP method could also be adapted to assign the presence of a specific mutation to the respective FUT3 alleles. We found the method to be reliable, rapid and cheap with no requirements for restriction enzyme processing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • DNA Mutational Analysis / methods
  • DNA Primers / genetics*
  • Diabetes Mellitus, Type 2 / enzymology
  • Diabetes Mellitus, Type 2 / genetics
  • Fucosyltransferases / genetics*
  • Genetic Testing / methods*
  • Genotype
  • Hemagglutination Tests
  • Humans
  • Mutation / genetics*
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Restriction Fragment Length
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Substrate Specificity
  • Time Factors

Substances

  • DNA Primers
  • Fucosyltransferases
  • 3-galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase
  • galactoside 2-alpha-L-fucosyltransferase