Primena nove generacije metoda za sekvenciranje dnk (next generation sequencing) u ranoj dijagnostici naslednih poremećaja

Abstract

U cilju rane dijagnostike i prevencije naslednih poremećaja, proteklih decenija su na raspolaganju bile različite metode. Analize genetičkog materijala su se kretale od klasične citogenetičke obrade kariotipa radi uočavanja numeričkih i strukturnih aberacija hromozoma, do najfinijih ispitivanja za detektuju genskih mutacija na molekularnom nivou. Poslednjih godina razvijaju se potpuno nove metode za brzu, efikasnu i dostupnu analizu naslednog materijala, koje su poznate kao “next generation sequencing” (NGS) ili nova generacija metoda za sekvenciranje DNK. Ove metode omogućavaju ispitivanje ne samo pojedinačnih gena ili delova gena nego i većeg broja segmenata, sve do kompletne nasledne osnove tj. čitavog genoma čoveka. Primena ovakvog pristupa dovodi do prave tihe revolucije u medicinskoj genetici i disciplinama sa kojima ona sarađuje, nagoveštavajući promenu u konceptu dijagnostike naslednih poremećaja. U prenatalnoj dijagnostici NGS je već našla primenu u potpuno neinvazivnoj detekciji najčešćih hromozomskih aberacija (Daunov, Edvardsov, Patau sindrom, aberacije polnih hromozoma) analizom fetalnih ćelija prisutnih u krvi majke. Test NIFTY već je dostupan i trudnicama u našoj sredini. U postnatalnom periodu NGS se koristi za ispitivanje odabranih panela gena, ili, po potrebi, čitavog genoma/ egzoma, sve sa ciljem što efikasnije dijagnostike pre svega monogenskih, ali i oligogenskih i poligenskih bolesti. Predlaže se čak da analiza kompletnog genoma postane deo neonatalnog skriniga, ali za sada to nije prihvaćeno. Nesumnjivo je da rezultati NGS donose veliki napredak medicinsko − genetičkoj praksi, ali i rađaju nove etičke dileme u oblasti rane detekcije naslednih poremećaja i intervencije kod ovih stanja.

In order to early diagnostics and prevention of hereditary disorders, past decades have been brought different methods. Analysis of genetic material ranged from classical cytogentic karyotype analysis for processing numerical and structural chromosomalaberratios, by most sophisticated examination of manor gene mutation on molecular level. In recent years develop completely new methods for rapid, efficient and publicly available analysis of inheritance material, that are known as the “next” button generation sequencing” (NGS). These methods allow for examination not only individual genes or parts of genes but also a larger number of segments, all up to complete inherited basis i.e. the entire human genome research. Implementation of this approach leads to genuine silent revolution in medical genetics and disciplines with which it cooperate, suggesting a change in the concept of heritage disorders diagnosing. In prenatal diagnostics NGS has already found application in completely noninvsive detection of chromosomal aberrations (Down, Edwards, Patau syndrome, sex chromosomes aberration) by analysis of fetal cells present in mother’s blood. Such tests (i.e. NIFTY) are already available and for pregnant women in our country. In postnatal period NGS is used for the examination of selected genes by gene panels, or, if necessary, the entire genome/exome research, all with the aim as efficient diagnostics primarily monogenic, but oligogenic and polygenic diseases also. It is proposed that the analysis of even complete genome become part of neonatal screening, but for now it is not accepted yet. It is undeniable that the results of NGS make a great progress in medical − genetic practice, but gained new ethical dilemmas in the field of early detection of hereditary disorders and intervention in these situations