medgenМедицинская генетикаMedical Genetics2073-7998Publishing House «Genius Media» LLC10.1234/XXXX-XXXX-2013-1-25-25medgen-21Research ArticleНАУЧНЫЕ ОБЗОРЫREVIEWТЕХНОЛОГИИ ВЫСОКОПАРАЛЛЕЛЬНОГО СЕКВЕНИРОВАНИЯ В МЕДИКО-ГЕНЕТИЧЕСКИХ ИССЛЕДОВАНИЯХNEXT GENERATION SEQUENCING TECHNOLOGIES IN MEDICAL GENETIC STUDIESТощаковС. В.ToshchakovS. V.

236041, Russia, Kaliningrad, A.Nevskogo str.

post@kantiana.ruДоминоваИ. Н.DominovaI. N.

236041, г.Калининград, ул. А.Невского, д.14

236041, Russia, Kaliningrad, A.Nevskogo str.

post@kantiana.ruПатрушевМ. В.PatrushevM. V.

236041, г.Калининград, ул. А.Невского, д.14

236041, Russia, Kaliningrad, A.Nevskogo str.

post@kantiana.ruФГАОУ ВПО «Балтийский федеральный университет им. И. Канта»РоссияImmanuel Kant Baltic Federal UniversityRussian Federation2013211220151211525Copyright © Тощаков С.В., Доминова И.Н., Патрушев М.В., 20152015Тощаков С.В., Доминова И.Н., Патрушев М.В.Toshchakov S.V., Dominova I.N., Patrushev 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/21

За последние 5 лет технологии секвенирования следующего поколения сыграли огромную роль в исследованиях генетических аспектов патогенеза различных заболеваний, и можно утверждать, что они стали мощнейшим толчком к появлению новой науки — медицинской геномики. Уже сейчас можно наблюдать многочисленные попытки внедрения высокопроизводительного секвенирования в клиническую диагностику, которые, однако, бессмысленны без глубокого понимания развития данных методик, их подводных камней и специфики. Задачей обзорной статьи являются детальное рассмотрение ныне существующих и активно разрабатываемых технологий высокопроизводительного секвенирования, а также обсуждение наиболее ярких примеров применения геномных технологий в медицине и потенциал их внедрения в рутинную практику медико-диагностических лабораторий.

In last 5 years next generation sequencing technologies made a huge impact on research of genetic aspects of pathogenesis and initiated the evolvement of the science of med ical genomics. At the moment we can observe multiple attempts of implementation of high-throughput sequencing in clinical diagnostics. Nevertheless those attempts might be meaningless without deep knowledge of such techniques, especially of its development, specificity and possible pitfalls. In this review we take a detailed overlook of existing and developing technologies and discuss most spectacular examples of its applications in medical genomics and its potential for implementation in routine diagnostic laboratory practice.

 

 

 

 

геномикасеквенированиесеквенирование следующего поколениянанопорыбиомаркёры мультифакториальных заболеванийgenomicssequencingnext generation sequenci ng technologiesnanoporesbiomarkers of multifactorial diseasesReferencesBrenner S. et al. Gene expression analysis by massively parallel signature sequencing (MPSS) on microbead arrays // Nature bio-techno-ogy. — 2000. — 18. — P. 630—634.Brenner S. et al. Gene expression analysis by massively parallel signature sequencing (MPSS) on microbead arrays // Nature bio-techno-ogy. — 2000. — 18. — P. 630—634.Chen W. et al. Mapping translocation breakpoints by next-generati-on sequencing // Genome research. — 2008. — 18. — P. 1143—1149.Chen W. et al. Mapping translocation breakpoints by next-generati-on sequencing // Genome research. — 2008. — 18. — P. 1143—1149.Flusberg B.A. et al. Direct detection of DNA methylation during single-molecule, real-time sequencing // Nature methods. — 2010. — 7. — P. 461—465.Flusberg B.A. et al. Direct detection of DNA methylation during single-molecule, real-time sequencing // Nature methods. — 2010. — 7. — P. 461—465.Garaj S. et al. Graphene as a subnanometre trans-electrode membrane // Nature. — 2010. — 467. — P. 190—193.Garaj S. et al. Graphene as a subnanometre trans-electrode membrane // Nature. — 2010. — 467. — P. 190—193.Hall A. R. et al. Hybrid pore formation by directed insertion of a-haemolysin into solid-state nanopores // Nature nanotechnology. — 2010. — 5. — P. 874—877.Hall A. R. et al. Hybrid pore formation by directed insertion of a-haemolysin into solid-state nanopores // Nature nanotechnology. — 2010. — 5. — P. 874—877.Harakalova M. et al. Genomic DNA pooling strategy for next-ge-neration sequencing-based rare variant discovery in abdominal aortic aneurysm regions of interest-challenges and limitations // Journal of cardiovascular translational research. — 2011. — 4. — P. 271—280.Harakalova M. et al. Genomic DNA pooling strategy for next-ge-neration sequencing-based rare variant discovery in abdominal aortic aneurysm regions of interest-challenges and limitations // Journal of cardiovascular translational research. — 2011. — 4. — P. 271—280.Howorka S., Cheley S., Bayley H. Sequence-specific detection of individual DNA strands using engineered nanopores // Nature biotechno-ogy. — 2001. — 19. — P. 636—639.Howorka S., Cheley S., Bayley H. Sequence-specific detection of individual DNA strands using engineered nanopores // Nature biotechno-ogy. — 2001. — 19. — P. 636—639.Kasianowicz J.J., Brandin E., Branton D., Deamer D.W. Characterization of individual polynucleotide molecules using a membrane channel // Proceedings of the National Academy of Sciences of the United States of America. — 1996. — 93. — P. 13770—13773.Kasianowicz J.J., Brandin E., Branton D., Deamer D.W. Characterization of individual polynucleotide molecules using a membrane channel // Proceedings of the National Academy of Sciences of the United States of America. — 1996. — 93. — P. 13770—13773.Ku C.S. et al. Technological advances in DNA sequence enrichment and sequencing for germline genetic diagnosis // Expert Rev. Mol. Diagn. — 2012. — 12(2). — P. 159—173.Ku C.S. et al. Technological advances in DNA sequence enrichment and sequencing for germline genetic diagnosis // Expert Rev. Mol. Diagn. — 2012. — 12(2). — P. 159—173.Lander E.S. et al. Initial sequencing and ana-ysis of the human genome // Nature. — 2001. — 409(6822). — P. 860—921.Lander E.S. et al. Initial sequencing and ana-ysis of the human genome // Nature. — 2001. — 409(6822). — P. 860—921.Lander E.S. Initial impact of the sequencing of the human genome // Nature. — 2011. — 470, — P. 187—197.Lander E.S. Initial impact of the sequencing of the human genome // Nature. — 2011. — 470, — P. 187—197.Levy S. et al. The diploid genome sequence of an individual human // PLoS biology. — 2007. — 5. — e254.Levy S. et al. The diploid genome sequence of an individual human // PLoS biology. — 2007. — 5. — e254.Luan B. et al. Base-by-base ratcheting of single stranded DNA through a solid-state nanopore // Physical review letters. — 2010. — 104, 238103.Luan B. et al. Base-by-base ratcheting of single stranded DNA through a solid-state nanopore // Physical review letters. — 2010. — 104, 238103.Lupski J.R. et al. Whole-genome sequencing in a patient with Charcot—Marie—Tooth neuropathy //The New England journal of medicine. — 2010. — 362. — P. 1181—1191.Lupski J.R. et al. Whole-genome sequencing in a patient with Charcot—Marie—Tooth neuropathy //The New England journal of medicine. — 2010. — 362. — P. 1181—1191.Margulies M. et al. Genome sequencing in microfabricated high-density picolitre reactors // Nature. — 2005. — 437. — P. 376—380.Margulies M. et al. Genome sequencing in microfabricated high-density picolitre reactors // Nature. — 2005. — 437. — P. 376—380.McNally B. et al. Optical recognition of converted DNA nucleotides for single-molecule DNA sequencing using nanopore arrays // Nano letters. — 2010. — 10. — P. 2237—2244.McNally B. et al. Optical recognition of converted DNA nucleotides for single-molecule DNA sequencing using nanopore arrays // Nano letters. — 2010. — 10. — P. 2237—2244.Meder B. et al. Targeted next-generation sequencing for the molecular genetic diagnostics of cardiomyopathies // Circulation. Cardiovascu-ar genetics. — 2011. — 4. — P. 110—122.Meder B. et al. Targeted next-generation sequencing for the molecular genetic diagnostics of cardiomyopathies // Circulation. Cardiovascu-ar genetics. — 2011. — 4. — P. 110—122.Michils G. et al. Mo-ecular analysis of the breast cancer genes BRCA1 and BRCA2 using amplicon-based massive parallel pyro-sequencing // J. Mol. Diagn. — 2012. — 14(6). — P. 623—630.Michils G. et al. Mo-ecular analysis of the breast cancer genes BRCA1 and BRCA2 using amplicon-based massive parallel pyro-sequencing // J. Mol. Diagn. — 2012. — 14(6). — P. 623—630.Nunnally B.K., He H., Li L.C., Tucker S.A., McGown L.B. Characterization of visible dyes for four-decay fluorescence detection in DNA sequencing // Ana-ytical chemistry. — 1997. — 69. — P. 2392—2397.Nunnally B.K., He H., Li L.C., Tucker S.A., McGown L.B. Characterization of visible dyes for four-decay fluorescence detection in DNA sequencing // Ana-ytical chemistry. — 1997. — 69. — P. 2392—2397.Ozcelik H. Long-range PCR and next-generation sequencing of BRCA1 and BRCA2 in breast cancer // J. Mol. Diagn. — 2012. — 14(5). — P. 467—475.Ozcelik H. Long-range PCR and next-generation sequencing of BRCA1 and BRCA2 in breast cancer // J. Mol. Diagn. — 2012. — 14(5). — P. 467—475.Parsons D.W. et al. An integrated genomic analysis of human glioblastoma multiforme // Science (New York, N.Y.). — 2008. — 321. — P. 1807—1812.Parsons D.W. et al. An integrated genomic analysis of human glioblastoma multiforme // Science (New York, N.Y.). — 2008. — 321. — P. 1807—1812.

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