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

• 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.

Ämnesord

MEDICIN OCH HÄLSOVETENSKAP  -- Klinisk medicin -- Annan klinisk medicin (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Clinical Medicine -- Other Clinical Medicine (hsv//eng)

Nyckelord

Alleles

DNA Mutational Analysis

methods

DNA Primers

genetics

Diabetes Mellitus

Type 2

enzymology

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

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)