A következő generációs szekvenálási technológiák megjelenése az elmúlt tíz évben jelentős előrelépést jelentett a gyors és hatékony genomiális DNS-szekvenálás terén. Tanulmányunkban áttekintjük az 1975-ös sangeri láncterminációs szekvenálástól a valós idejű DNS-szekvenálás lehetővé válásáig vezető módszertani vívmányokat. A klonális amplikonokkal dolgozó, sok szálon párhuzamosan futó szekvenálási módszerek a következő generációs szekvenálási technológiák alapjai. Manapság leginkább a funkcionális genomikai alapkutatásban alkalmazzák ezen szekvenálási technológiákat, amelyek a szignáltranszdukciós útvonalak, ontológiák, a proteomikai, metabolomikai eredményeknek a metaanalízise során nélkülözhetetlen információt adnak. Bár klinikumban rutinmódon még csak elvétve alkalmaznak következő generációs szekvenátorokat, azonban az onkológiában, kardiológiában és epidemiológiában már van igény a technológia által elérhető extra ismeretekre. Az elterjedés fő gátja az adatelemzési módszerek standardizáltságnak hiánya, amely az objektív kiértékelést megnehezíti. Orv. Hetil., 2011, 152, 55–62.
Lander, E. S., Linton, L. M., Birren, B. és mtsai: Initial sequencing and analysis of the human genome. Nature, 2001, 409, 860–921.
Birren B. , 'Initial sequencing and analysis of the human genome ' (2001 ) 409 Nature : 860 -921 .
Venter, J. C., Adams, M. D., Myers, E. W. és mtsai: The sequence of the human genome. Science, 2001, 291, 1304–1351.
Myers E. W. , 'The sequence of the human genome ' (2001 ) 291 Science : 1304 -1351 .
Sanger, F.: Nucleotide sequences in DNA. Proc. R. Soc. Lond B. Biol. Sci., 1975, 191, 317–333.
Sanger F. , 'Nucleotide sequences in DNA ' (1975 ) 191 Proc. R. Soc. Lond B. Biol. Sci. : 317 -333 .
Smith, L., Sanders, J., Kaiser, R. és mtsai: Fluorescence detection in automated DNA sequence analysis. Nature, 1986, 321, 674–679.
Kaiser R. , 'Fluorescence detection in automated DNA sequence analysis ' (1986 ) 321 Nature : 674 -679 .
Metzker, M. L., Raghavachari, R., Richards, S. és mtsai: Termination of DNA synthesis by novel 3’-modified-deoxyribonucleoside 5’-riphosphates. Nucleic Acids Res., 1994, 22, 4259–4267.
Richards S. , 'Termination of DNA synthesis by novel 3’-modified-deoxyribonucleoside 5’-riphosphates ' (1994 ) 22 Nucleic Acids Res. : 4259 -4267 .
Ruiz-Martinez, M. C., Berka, J., Belenkii, A. és mtsai: DNA sequencing by capillary electrophoresis with replaceable linear polyacrylamide and laser-induced fluorescence detection. Anal. Chem., 1993, 65, 2851–2858.
Belenkii A. , 'DNA sequencing by capillary electrophoresis with replaceable linear polyacrylamide and laser-induced fluorescence detection ' (1993 ) 65 Anal. Chem. : 2851 -2858 .
Ju, J., Ruan, C., Fuller, C. és mtsa: Fluorescence energy transfer dye-labeled primers for DNA sequencing and analysis. Proc. Natl. Acad. Sci., 1995, 92, 4347–4351.
Fuller C. , 'Fluorescence energy transfer dye-labeled primers for DNA sequencing and analysis ' (1995 ) 92 Proc. Natl. Acad. Sci. : 4347 -4351 .
Lee, L., Spurgeon, S., Heiner, C. és mtsai: New energy transfer dyes for DNA sequencing. Nucleic Acids Res., 1997, 25, 2816–2822.
Heiner C. , 'New energy transfer dyes for DNA sequencing ' (1997 ) 25 Nucleic Acids Res. : 2816 -2822 .
Kheterpal, I., Scherer, J., Clark, S. és mtsai: DNA sequencing using a four-color confocal fluorescence capillary array scanner. Electrophoresis, 1996, 17, 1852–1859.
Clark S. , 'DNA sequencing using a four-color confocal fluorescence capillary array scanner ' (1996 ) 17 Electrophoresis : 1852 -1859 .
Ewing, B., Green, P.: Base-calling of automated sequencer traces using Phred. II. Error probabilities. Genome Res., 1998, 8, 186–194.
Green P. , 'Base-calling of automated sequencer traces using Phred. II. Error probabilities ' (1998 ) 8 Genome Res. : 186 -194 .
Ewing, B., Hillier, L., Wendl, M. C. és mtsa: Base-calling of automated sequencer traces using Phred. I. Accuracy assessment. Genome Res., 1998, 8, 175–185.
Wendl M. C. , 'Base-calling of automated sequencer traces using Phred. I. Accuracy assessment ' (1998 ) 8 Genome Res. : 175 -185 .
Schmalzing, D., Tsao, N., Koutny, L. és mtsai: Toward real-world sequencing by microdevice electrophoresis. Genome Res., 1999, 9, 853–858.
Koutny L. , 'Toward real-world sequencing by microdevice electrophoresis ' (1999 ) 9 Genome Res. : 853 -858 .
Liu, S., Shi, Y., Ja, W. és mtsa: Optimization of high-speed DNA sequencing on microfabricated capillary electrophoresis channels. Anal. Chem, 1999, 71, 566–573.
Ja W. , 'Optimization of high-speed DNA sequencing on microfabricated capillary electrophoresis channels ' (1999 ) 71 Anal. Chem : 566 -573 .
Liu, S., Ren, H., Gao, Q. és mtsai: Automated parallel DNA sequencing on multiple channel microchips. Proc. Natl. Acad. Sci., 2000, 97, 5369–5374.
Gao Q. , 'Automated parallel DNA sequencing on multiple channel microchips ' (2000 ) 97 Proc. Natl. Acad. Sci. : 5369 -5374 .
Simpson, J. W., Ruiz-Martinez, M. C., Mulhern, G. T. és mtsai: Transmission imaging spectrograph and microfabricated channel system for DNA analysis. Electrophoresis, 2000, 21, 135–149.
Mulhern G. T. , 'Transmission imaging spectrograph and microfabricated channel system for DNA analysis ' (2000 ) 21 Electrophoresis : 135 -149 .
Backhouse, C., Caamano, M., Oaks, F. és mtsai: DNA sequencing in a monolithic microchannel device. Electrophoresis, 2000, 21, 150–156.
Oaks F. , 'DNA sequencing in a monolithic microchannel device ' (2000 ) 21 Electrophoresis : 150 -156 .
Koutny, L., Schmalzing, D., Salas-Solano, O. és mtsai: Eight hundred-base sequencing in a microfabricated electrophoretic device. Anal. Chem., 2000, 72, 3388–3391.
Salas-Solano O. , 'Eight hundred-base sequencing in a microfabricated electrophoretic device ' (2000 ) 72 Anal. Chem. : 3388 -3391 .
Lewis, E. K., Haaland, W. C., Nguyen, F. és mtsai: Color-blind fluorescence detection for four-color DNA sequencing. Proc. Natl. Acad. Sci., 2005, 102, 5346–5351.
Nguyen F. , 'Color-blind fluorescence detection for four-color DNA sequencing ' (2005 ) 102 Proc. Natl. Acad. Sci. : 5346 -5351 .
Emrich, C. A., Tian, H., Medintz, I. L. és mtsa: Microfabricated 384-lane capillary array electrophoresis bioanalyzer for ultrahigh-throughput genetic analysis. Anal. Chem, 2002, 74, 5076–5083.
Medintz I. L. , 'Microfabricated 384-lane capillary array electrophoresis bioanalyzer for ultrahigh-throughput genetic analysis ' (2002 ) 74 Anal. Chem : 5076 -5083 .
Pettersson, E., Lundeberg, J., Ahmadian, A.: Generations of sequencing technologies. Genomics, 2009, 93, 105–111.
Ahmadian A. , 'Generations of sequencing technologies ' (2009 ) 93 Genomics : 105 -111 .
Mardis, E. R.: The impact of next-generation sequencing technology on genetics. Trends Genet., 2008, 24, 133–141.
Mardis E. R. , 'The impact of next-generation sequencing technology on genetics ' (2008 ) 24 Trends Genet. : 133 -141 .
Margulies, M., Egholm, M., Altman, W. E. és mtsai: Genome sequencing in microfabricated high-density picolitre reactors. Nature, 2005, 437, 376–380.
Altman W. E. , 'Genome sequencing in microfabricated high-density picolitre reactors ' (2005 ) 437 Nature : 376 -380 .
Ronaghi, M., Uhlen, M., Nyren, P.: A sequencing method based on real-time pyrophosphate. Science, 1998, 281, 363–365.
Nyren P. , 'A sequencing method based on real-time pyrophosphate ' (1998 ) 281 Science : 363 -365 .
Korbel, J. O., Urban, A. E., Affourtit, J. P. és mtsai: Paired-end mapping reveals extensive structural variation in the human genome. Science, 2007, 318, 420–426.
Affourtit J. P. , 'Paired-end mapping reveals extensive structural variation in the human genome ' (2007 ) 318 Science : 420 -426 .
Harris, T. D., Buzby, P. R., Babcock, H. és mtsai: Single-molecule DNA sequencing of a viral genome. Science, 2008, 320, 106–109.
Babcock H. , 'Single-molecule DNA sequencing of a viral genome ' (2008 ) 320 Science : 106 -109 .
Eid, J., Fehr, A., Gray, J. és mtsai: Real-time DNA sequencing from single polymerase molecules. Science, 2009, 323, 133–138.
Gray J. , 'Real-time DNA sequencing from single polymerase molecules ' (2009 ) 323 Science : 133 -138 .
Fologea, D., Gershow, M., Ledden, B. és mtsai: Detecting single stranded DNA with a solid state nanopore. Nano Lett., 2005, 10, 1905–1909.
Ledden B. , 'Detecting single stranded DNA with a solid state nanopore ' (2005 ) 10 Nano Lett. : 1905 -1909 .
Stoddart, D., Maglia, G., Mikhailova, E. és mtsa: Multiple base-recognition sites in a biological nanopore: two heads are better than one. Angew. Chem. Int. Ed. Engl., 2010, 49, 556–559.
Mikhailova E. , 'Multiple base-recognition sites in a biological nanopore: two heads are better than one ' (2010 ) 49 Angew. Chem. Int. Ed. Engl. : 556 -559 .
Varki, A., Geschwind, D. H., Eichler, E. E.: Explaining human uniqueness: genome interactions with environment, behaviour and culture. Nat. Rev. Genet., 2008, 9, 749–763.
Eichler E. E. , 'Explaining human uniqueness: genome interactions with environment, behaviour and culture ' (2008 ) 9 Nat. Rev. Genet. : 749 -763 .
Beyer, A., Bandyopadhyay, S., Ideker, T.: Integrating physical and genetic maps: from genomes to interaction networks. Nat. Rev. Genet., 2007, 8, 699–710.
Ideker T. , 'Integrating physical and genetic maps: from genomes to interaction networks ' (2007 ) 8 Nat. Rev. Genet. : 699 -710 .
Hu, Z., Killion, P. J., Iyer, V. R.: Genetic reconstruction of a functional transcriptional regulatory network. Nat. Genet., 2007, 39, 683–687.
Iyer V. R. , 'Genetic reconstruction of a functional transcriptional regulatory network ' (2007 ) 39 Nat. Genet. : 683 -687 .
Werner, T.: Regulatory networks: linking microarray data to systems biology. Mech. Ageing Dev., 2007, 128, 168–172.
Werner T. , 'Regulatory networks: linking microarray data to systems biology ' (2007 ) 128 Mech. Ageing Dev. : 168 -172 .
Thomas, P. D., Mi, H., Lewis, S.: Ontology annotation: mapping genomic regions to biological function. Curr. Opin. Chem. Biol., 2007, 11, 4–11.
Lewis S. , 'Ontology annotation: mapping genomic regions to biological function ' (2007 ) 11 Curr. Opin. Chem. Biol. : 4 -11 .
Werner, T.: Bioinformatics applications for pathway analysis of microarray data. Curr. Opin. Biotechnol., 2008, 19, 50–54.
Werner T. , 'Bioinformatics applications for pathway analysis of microarray data ' (2008 ) 19 Curr. Opin. Biotechnol. : 50 -54 .
Kertesz, M., Wan, Y., Mazor, E. és mtsai: Genome-wide measurement of RNA secondary structure in yeast. Nature, 2010, 467, 103–107.
Mazor E. , 'Genome-wide measurement of RNA secondary structure in yeast ' (2010 ) 467 Nature : 103 -107 .
Werner, T.: Next generation sequencing in functional genomics. Brief Bioinform., 2010, 5, 449–511.
Werner T. , 'Next generation sequencing in functional genomics ' (2010 ) 5 Brief Bioinform. : 449 -511 .
Kumar, M., Tanwar, M., Saxena, R. és mtsa: Identification of novel mitochondrial mutations in Leber’s hereditary optic neuropathy. Mol. Vis., 2010, 16, 782–792.
Saxena R. , 'Identification of novel mitochondrial mutations in Leber’s hereditary optic neuropathy ' (2010 ) 16 Mol. Vis. : 782 -792 .
Lewis, T., Loman, N. J., Bingle, L. és mtsai: High-throughput whole-genome sequencing to dissect the epidemiology of Acinetobacter baumannii isolates from a hospital outbreak. J. Hosp. Infect., 2010, 75, 37–41.
Bingle L. , 'High-throughput whole-genome sequencing to dissect the epidemiology of Acinetobacter baumannii isolates from a hospital outbreak ' (2010 ) 75 J. Hosp. Infect. : 37 -41 .
Fokstuen, S., Lyle, R., Munoz, A. és mtsai: A DNA resequencing array for pathogenic mutation detection in hypertrophic cardiomyopathy. Hum. Mutat., 2008, 29, 879–885.
Munoz A. , 'A DNA resequencing array for pathogenic mutation detection in hypertrophic cardiomyopathy ' (2008 ) 29 Hum. Mutat. : 879 -885 .
Fan, H. C., Blumenfeld, Y. J., Chitkara, U. és mtsai: Noninvasive diagnosis of fetal aneuploidy by shotgun sequencing DNA from maternal blood. Proc. Natl. Acad. Sci. USA, 2008, 105, 16266–16271.
Chitkara U. , 'Noninvasive diagnosis of fetal aneuploidy by shotgun sequencing DNA from maternal blood ' (2008 ) 105 Proc. Natl. Acad. Sci. USA : 16266 -16271 .
Chiu, R. W., Chan, K. C., Gao, Y. és mtsai: Noninvasive prenatal diagnosis of fetal chromosomal aneuploidy by massively parallel genomic sequencing of DNA in maternal plasma. Proc. Natl. Acad. Sci. USA, 2008, 105, 20458–20463.
Gao Y. , 'Noninvasive prenatal diagnosis of fetal chromosomal aneuploidy by massively parallel genomic sequencing of DNA in maternal plasma ' (2008 ) 105 Proc. Natl. Acad. Sci. USA : 20458 -20463 .
Papadopoulos, N., Kinzler, K. W., Vogelstein, B.: The role of companion diagnostics in the development and use of mutation-targeted cancer therapies. Nat. Biotechnol., 2006, 8, 985–995.
Vogelstein B. , 'The role of companion diagnostics in the development and use of mutation-targeted cancer therapies ' (2006 ) 8 Nat. Biotechnol. : 985 -995 .
Ding, L., Getz, G., Wheeler, D. A. és mtsai: Somatic mutations affect key pathways in lung adenocarcinoma. Nature, 2008, 455, 1069–1075.
Wheeler D. A. , 'Somatic mutations affect key pathways in lung adenocarcinoma ' (2008 ) 455 Nature : 1069 -1075 .
Greenman, C., Stephens, P., Smith, R. és mtsai: Patterns of somatic mutation in human cancer genomes. Nature, 2007, 446, 153–158.
Smith R. , 'Patterns of somatic mutation in human cancer genomes ' (2007 ) 446 Nature : 153 -158 .
Davies, H., Bignell, G. R., Cox C. és mtsai: Mutations of the BRAF gene in human cancer. Nature, 2002, 417, 949–954.
Cox C. , 'Mutations of the BRAF gene in human cancer ' (2002 ) 417 Nature : 949 -954 .
Sharma, S. V., Bell, D. W., Settleman, J. és mtsa: Epidermal growth factor receptor mutations in lung cancer. Nat. Rev. Cancer, 2007, 7, 169–181.
Settleman J. , 'Epidermal growth factor receptor mutations in lung cancer ' (2007 ) 7 Nat. Rev. Cancer : 169 -181 .
Carpten, J. D., Faber, A. L., Horn, C. és mtsai: A transforming mutation in the pleckstrin homology domain of AKT1 in cancer. Nature, 2007, 448, 439–444.
Horn C. , 'A transforming mutation in the pleckstrin homology domain of AKT1 in cancer ' (2007 ) 448 Nature : 439 -444 .
Samuels, Y., Wang, Z., Bardelli, A. és mtsai: High frequency of mutations of the PIK3CA gene in human cancers. Science, 2004, 304, 554.
Bardelli A. , 'High frequency of mutations of the PIK3CA gene in human cancers ' (2004 ) 304 Science : 554 -.
Kohlmann, A., Grossmann, V., Klein, H. U. és mstai: Next-generation sequencing technology reveals a characteristic pattern of molecular mutations in 72.8% of chronic myelomonocytic leukemia by detecting frequent alterations in TET2, CBL, RAS, and RUNX1. J. Clin. Oncol., 2010, 28, 3858–3865.
Klein H. U. , 'Next-generation sequencing technology reveals a characteristic pattern of molecular mutations in 72.8% of chronic myelomonocytic leukemia by detecting frequent alterations in TET2, CBL, RAS, and RUNX1 ' (2010 ) 28 J. Clin. Oncol. : 3858 -3865 .
Pfeifer, G. P., Besaratinia, A.: Mutational spectra of human cancer. Hum. Genet., 2009, 125, 493–506.
Besaratinia A. , 'Mutational spectra of human cancer ' (2009 ) 125 Hum. Genet. : 493 -506 .