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Development of a high-throughput fluorescence assay for detecting SNPs in hemostasis and folate metabolism genes for clinical use

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Abstract

The genetic predisposition of an individual patient should be taken into account to choose the proper treatment. Doing this in clinical practice requires the development of rapid, high-throughput, and easy-to-do assays intended to detect single nucleotide polymorphisms. A detection kit intended to identify the hemostasis and folate cycle gene mutations G20210A FII, G1691A FV, G10976A FVII, G103T FXIII, C807T ITGA2, T1565C ITGB3, 5G(-675)4G PAI, G(-455)A FGB, C677T and A1298C MTHFR, A2756G MTR, and A66G MTRR is proposed in this work. The method is based on the polymerase chain reaction and subsequent melt curve analysis of complexes of amplicons with a specific probe. Three single-nucleotide polymorphisms can be identified in one tube using our detection kit, which increases the productivity of the analysis in clinical use. Different types of biological samples (buccal epithelium, saliva, plasma, serum, and urogenital swabs) can be used as the initial material for DNA isolation and further analysis by the method developed in the work.

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Authors and Affiliations

  1. Institute of Cytology and Genetics, Siberian Branch, Russian Academy of Sciences, pr. Akademika Lavrent’eva 10, Novosibirsk, Russia

    M. A. Prasolova & G. M. Dymshits

  2. JSC Vector-Best, ABK, Koltsovo, Novosibirsk region, 630559, Russia

    M. A. Prasolova & E. G. Shchepotina

  3. Department of Molecular Biology, Novosibirsk State University, ul. Pirogova 2, Novosibirsk, 630090, Russia

    G. M. Dymshits

Authors
  1. M. A. Prasolova
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  2. E. G. Shchepotina
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  3. G. M. Dymshits
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Corresponding author

Correspondence to M. A. Prasolova.

Additional information

Original Russian Text © M.A. Prasolova, E.G. Shchepotina, G.M. Dymshits, 2013, published in Molekulyarnaya Genetika, Mikrobiologiya i Virusologiya, 2013, No. 1, pp. 23–29.

The article was translated by the authors.

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Prasolova, M.A., Shchepotina, E.G. & Dymshits, G.M. Development of a high-throughput fluorescence assay for detecting SNPs in hemostasis and folate metabolism genes for clinical use. Mol. Genet. Microbiol. Virol. 28, 24–31 (2013). https://doi.org/10.3103/S0891416813010047

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  • DOI: https://doi.org/10.3103/S0891416813010047

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