Simultaneous Genotyping of Four Single Nucleotide Polymorphisms Associated with Risk Factors of Hemostasis Disorders

Author(s): Eugenia E. Bashmakova, Vasilisa V. Krasitskaya, Ludmila A. Frank

Journal Name: Combinatorial Chemistry & High Throughput Screening
Accelerated Technologies for Biotechnology, Bioassays, Medicinal Chemistry and Natural Products Research

Volume 18 , Issue 10 , 2015


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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.

Keywords: SNP detection, PEXT reaction, photoprotein obelin, bioluminescent microassay, multiplex PCR.

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Article Details

VOLUME: 18
ISSUE: 10
Year: 2015
Published on: 16 September, 2015
Page: [930 - 936]
Pages: 7
DOI: 10.2174/1386207318666150917100134
Price: $65

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