medgenМедицинская генетикаMedical Genetics2073-7998Publishing House «Genius Media» LLCmedgen-344Research ArticleОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯORIGINAL RESEARCHРазработка технологии диагностики наследственных генетических вариантов, определяющих чувствительность к варфарину, методом однонуклеотидного удлинения праймеровThe developing of technology for diagnostics of genetic variants, that determine the sensitivity to warfarin, using the single nucleotide primer extension methodБабушкинаН. П.BabushkinaN. P.nad.babushkina@medgenetics.ruГончароваИ. А.GoncharovaI. A.noemail@neicon.ruГолубенкоМ. В.GolubenkoM. V.noemail@neicon.ru«Томский национальный исследовательский медицинский центр Российской академии наук» Научно-исследовательский институт медицинской генетикиРоссияResearch Institute of Medical Genetics, Tomsk NRMCRussian Federation20172002201816112731Copyright © Бабушкина Н.П., Гончарова И.А., Голубенко М.В., 20182018Бабушкина Н.П., Гончарова И.А., Голубенко М.В.Babushkina N.P., Goncharova I.A., Golubenko M.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.medgen-journal.ru/jour/article/view/344

Варфарин назначается большому количеству пациентов при различных заболеваниях, однако существуют 20-кратные межиндивидуальные различия требуемой дозировки; превышение же необходимой дозы чревато осложнениями в виде кровотечений различной локализации, вплоть до развития жизнеугрожающих состояний. Известны генетические факторы, оказывающие существенное влияние на метаболизм варфарина в организме. Цель настоящего исследования заключалась в разработке набора олигонуклеотидов и методики генотипирования генетических полиморфизмов, влияющих на эффективность действия варфарина. Разработанная мультиплексная панель включает 6 полиморфных вариантов в генах биотрансформации, сочетания генотипов которых определяют подбор правильной дозы варфарина: CYP2C9 (rs1799853, rs1057910), VKORC1 (rs9923231, rs8050894), CYP4F2 (rs2108622) и GGCX (rs11676382). Для разработки набора проб были проанализированы тип генотипируемых замен, и характеристики ДНК в регионе данных SNP; возможность мультиплексирования проверяли на основании результатов реакции с набором Primer Focus Kit. Проведено мультиплексное генотипирование контрольных образцов ДНК с известными генотипами. Генотипы, определенные методом SNaPshot-анализа соответствовали полученным стандартными методами. Таким образом, разработана и верифицирована панель проб для мультиплексного генотипирования шести SNP, связанных с индивидуальной чувствительностью к варфарину. Использование в клинической практике результатов данного анализа позволит повысить эффективность антикоагулянтной терапии и снизить частоту проявления побочных эффектов.

Warfarin is prescribed to a lot of patients with different pathologies, however there are individual differences of required dosage up to twenty times. Overdosage can cause health complications like different localized bleeding, up to if threatening condition. There are genetics factors, that influences on warfarin metabolism in the human body. The point of this research is the development of set of oligonucleotide and genotyping of genetic polymorphism, that influence on warfarin effect. The developed multiplex panel (мультиплексная панель) includes six polymorphic variants in biotransformation genes, and their combination determine correct warfarin dose: CYP2C9 (rs1799853, rs1057910), VKORC1 (rs9923231, rs8050894), CYP4F2 (rs2108622) and GGCX (rs11676382).Genetic replacement types and DNA characteristics in SNP region were analysed to develop the sample set. The multiplexing opportunity was checked according to the results of the reaction with Primer Focus Kit. Multiplex genotyping of DNA control samples with known genotypes was performed. Genotypes wich were identified using SNaPshot, corresponds to those, indentified using standard method. In summary, sample panel for multiplex genotyping of six SNP was developed and verified, SNP are connected with individual warfarin sensibility. Using these results in clinical practice can increase efficiency of anticoagulant therapy and decrease side effects frequency.

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The authors declare that there are no conflicts of interest present.