vestichИзвестия Национальной академии наук Беларуси. Серия химических наукProceedings of the National Academy of Sciences of Belarus, Chemical Series1561-83312524-2342The Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"10.29235/1561-8331-2021-57-1-48-60vestich-631Research ArticleБИООРГАНИЧЕСКАЯ ХИМИЯBIOORGANIC CHEMISTRYРазработка мультиплексной системы для определения маркеров предрасположенности к развитию сердечно-сосудистых заболеванийDevelopment of the multiplex system for determining markers of predisposition to cardiovascular diseasesГайдукевичИ. В.HaidukevichI. V.

Гайдукевич Ирина Витальевна - кандидат химических наук, старший научный сотрудник.ул. Акад. В.Ф. Купревича, 5/2, 220141, Минск.

Irina V. Haidukevich - Ph. D. (Chemistry), Senior Researcher. Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus.5/2, Acad. Kuprevich Str., Minsk.

ribano4ka@gmail.comГорькаваяА. М.GorkavayaA. M.

Горькавая Анна Михайловна - научный сотрудник.ул. Акад. В.Ф. Купревича, 5/2, 220141, Минск.

Anna M. Gorkavaya - Researcher, Institute of Bioor-ganic Chemistry of the National Academy of Sciences of Belarus.5/2, Acad. Kuprevich Str., Minsk.

annagorkavaya@gmail.comГрудоА. В.GrudoA. V.

Грудо Анна Викторовна - кандидат химических наук, старший научный сотрудник.ул. Акад. В.Ф. Купревича, 5/2, 220141, Минск.

Anna V. Grudo - Ph. D. (Chemistry), Senior Researcher. Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus.5/2, Acad. Kuprevich Str., Minsk.

vasilevskaya.av@gmail.comСергеевГ. В.SergeevG. V.

Сергеев Геннадий Валерьевич - кандидат химических наук, заведующий лабораторией.ул. Акад. В.Ф. Купревича, 5/2, 220141, Минск.

Gennady V. Sergeev - Ph. D. (Chemistry), Head of the laboratory. Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus.5/2, Acad. Kuprevich Str., Minsk.

gvserg@iboch.byУсановС. А.UsanovS. A.

Усанов Сергей Александрович - член-корреспондент, доктор химических наук, профессор, главный научный сотрудник.ул. Акад. В.Ф. Купревича, 5/2, 220141, Минск.

Sergei A. Usanov - Corresponding Member of the National Academy of Sciences of Belarus, D. Sc. (Chemistry), Professor, Principal Researcher. Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus.5/2, Acad. Kuprevich Str., Minsk.

usanov@iboch.byИнститут биоорганической химии Национальной академии наук БеларусиInstitute of Bioorganic Chemistry of the National Academy of Sciences of Belarus2021090220215714860Copyright © Гайдукевич И.В., Горькавая А.М., Грудо А.В., Сергеев Г.В., Усанов С.А., 20212021Гайдукевич И.В., Горькавая А.М., Грудо А.В., Сергеев Г.В., Усанов С.А.Haidukevich I.V., Gorkavaya A.M., Grudo A.V., Sergeev G.V., Usanov S.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestichem.belnauka.by/jour/article/view/631

Разработана мультиплексная система для определения мутаций в локусах rs5985, rs1799983, rs5918, rs2243093, rs4673, rs4646994, rs1722009, rs3980933, rs71103505, ассоциированных с развитием сердечно-сосудистых заболеваний. Данные мутации относятся к различным типам - SNP, STR, Ins/Del - поэтому для их определения использованы технологии минисеквенирования и фрагментного анализа. Проведен анализ олигонуклеотидов для амплификации всех локусов в формате одной реакции. Технология минисеквенирования по сравнению с фрагментным анализом потребовала дополнительных этапов пробоподготовки, в связи с чем олигонуклеотиды для локусов с SNP были объединены в отдельный плекс. Для двух созданных плексов оптимизированы одинаковые состав амплификационной смеси и параметры программы ПЦР реакции, разработаны панели «бинов» для интерпретации результатов. Апробация системы подтвердила возможность определения мутаций в девяти локусах с высокой чувствительностью и воспроизводимостью.

A multiplex system to detect mutations at loci rs5985, rs1799983, rs5918, rs2243093, rs4673, rs4646994, rs1722009, rs3980933, rs71103505 associated with the development of cardiovascular diseases has been developed. These mutations belong to different types - SNP, STR, Ins/Del - therefore, minisequencing and fragment analysis technologies were used to determine them. Oligonucleotide analysis was performed to amplify all loci in a single reaction format. The minisequencing technology, in comparison with fragment analysis, required additional stages of sample preparation; therefore, oligonucleotides for loci with SNP were combined into a separate plex. For the two plexes created, the same composition of the amplification mixture and parameters of the PCR reaction program were optimized, and “bin” panels were developed to interpret the results. The testing of the system confirmed the possibility of detecting mutations at nine loci with high sensitivity and reproducibility.

 

фрагментный анализминисеквенированиеSNPсердечно-сосудистые заболеванияfragment analysisminisequencingSNPcardiovascular diseaseФинансирование работ осуществлялось в рамках Мероприятия 9 НТП Союзного государства «ДНК-идентификация». Авторы выражают благодарность за предоставление информации о маркерах, ассоциированных с развитием сердечно-сосудистых заболеваний, сотрудникам А.Л. Гончару и М.Д. Амельяновичу Института генетики и цитологии НАН Беларуси.Funding for the work was carried out within the framework of Event 9 of the Scientific and Technical Programm of the Union State “DNA identification”. The authors are grateful to the staff of the Institute of Genetics and Cytology of the National Academy of Sciences of Belarus A.L. Gonchar and M.D. Amelyanovich for providing information on markers associated with the development of cardiovascular diseases.ReferencesGenome-wide polygenic scores for common diseases identify individuals with risk equivalent to monogenic mutations / A. V. 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The authors declare that there are no conflicts of interest present.