Title:Simultaneous Genotyping of Four Single Nucleotide Polymorphisms Associated with Risk Factors of Hemostasis Disorders
Volume: 18
Issue: 10
Author(s): Eugenia E. Bashmakova, Vasilisa V. Krasitskaya and Ludmila A. Frank
Affiliation:
Keywords:
SNP detection, PEXT reaction, photoprotein obelin, bioluminescent microassay, multiplex PCR.
Abstract: Multiplex simultaneous genotyping technique was developed for four polymorphisms in
genes coding for blood coagulation factors and homocysteine metabolism which are considered as
thrombophilia related mutations: FV Leiden, FII G20210A, MTHFR C677T, and FVII G10976A. It is
based on primer extension reaction with the following bioluminescent solid-phase microassay. At that,
two dierent in bioluminescence obelin mutants were applied to simultaneous detection of two gene
allelic variants. The assay is carried out in microtiter plate format and provides fast and reliable genotyping of four single
nucleotide polymorphisms in four different genes within 2.5 hours. A large number of clinical samples were analyzed and
the obtained results were found to be in complete correlation with those obtained by using conventional RT-PCR
techniques.
