Determination of Lewis FUT3 gene mutations by PCR using sequence-specific primers enables efficient genotyping of clinical samples
✍ Scribed by Ammi Grahn; Anders Elmgren; Lena Åberg; Lola Svensson; Per-Anders Jansson; Peter Lönnroth; Göran Larson
- Publisher
- John Wiley and Sons
- Year
- 2001
- Tongue
- English
- Weight
- 69 KB
- Volume
- 18
- Category
- Article
- ISSN
- 1059-7794
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✦ Synopsis
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.