izpis_h1_title_alt

Opredelitev kriterijev za razširjeno presejanje novorojencev za vrojene bolezni presnove : doktorska disertacija
ID Šmon, Andraž (Author), ID Trebušak Podkrajšek, Katarina (Mentor) More about this mentor... This link opens in a new window, ID Marc, Janja (Co-mentor)

.pdfPDF - Presentation file, Download (2,26 MB)
MD5: F90C1B56F35F2E9B4D9CF51B81B6E7F8

Abstract
Vrojene bolezni presnove (VBP) so redke bolezni, večina jih ima pojavnost, manjšo od 1 : 10.000. Zgodnje odkrivanje VBP je ključno za pravilne in pravočasne klinične odločitve, ki lahko preprečijo hude posledice bolezni ali celo smrt. V nasprotju s številnimi evropskimi državami v državah jugovzhodne Evrope ni vpeljano razširjeno presejanje novorojencev (NBS) s tandemsko masno spektrometrijo, temveč poteka presejanje samo za nekaj bolezni večinoma z uporabo imunokemijskih in fluorimetričnih metod. Namen dela je bil pripraviti optimalno strategijo za organiziranje razširjenega NBS za VBP v Sloveniji, ki temelji na izbranih kriterijih in izdelanem točkovalniku. Kriterije in točkovalnik smo oblikovali in ovrednotili na osnovi eksperimentalnih rezultatov naše raziskave in na ta način določili nabor bolezni, ki bi ga bilo smiselno vključiti v program razširjenega NBS za slovensko populacijo. Izvedli smo retrospektivno raziskavo, v kateri smo analizirali vzorce posušenih kapelj krvi na filtrirnem papirju 10.048 preiskovancev. V začetni stopnji presejanja smo s tandemsko masno spektrometrijo posamezno VBP prepoznali ter jo v nadaljevanju s potrditveni testi potrdili. Pri tem smo uporabili tudi metodo sekvenciranja naslednje generacije (NGS). Ta metoda omogoča sočasno analizo vseh izbranih genov, povezanih z VBP, in tako predstavlja metodo izbire za potrjevanje bolezni, prepoznanih pri NBS. Zaradi nejasnih rezultatov NBS smo naknadno 113 preiskovancev povabili na kontrolni pregled in dodatne potrditvene analize. Potrditveni testi so vključevali analizo posušenih kapelj krvi na filtrirnem papirju, analizo organskih kislin v urinu, analizo aminokislin v plazmi ter v izbranih primerih tudi analizo encimske aktivnosti v fibroblastih ter acilkarnitinsko profiliranje v fibroblastih. Na osnovi rezultatov analize 10.048 preiskovancev in primerjave z evropskimi državami smo opredelili izključitvene vrednosti za posamezne presnovke in razmerja med njimi ter razvili sistem točkovanja, ki je omogočal, da smo za posamezno VBP lahko prepoznali preiskovance z najvišjim tveganjem za razvoj VBP. Na osnovi tega smo pripravili predlog nabora bolezni, ki bi jih bilo smotrno vključiti v program razširjenega NBS v Sloveniji. Na novo smo s pomočjo potrditvenih testov odkrili štiri bolnike z VBP, in sicer bolnika z glutarično acidurijo tipa 1, bolnika s pomanjkanjem zelo-dolgoverižne-acil-CoAdehidrogenaze in dva bolnika s pomanjkanjem 3-metilkrotonil-CoA-karboksilaze. Pri vseh smo s pristopom NGS opredelili vzročne genetske spremembe, med njimi takšne, ki še niso bile opisane v literaturi. Še neopisani genetski spremembi sta bili vzročna genetska sprememba c.205-7-8delCTinsGC v genu ACADVL ter benigna genetska sprememba c.149_151dupCCA (p.Thr50dup) v genu MCCC1. S pomočjo rezultatov naše raziskave in na osnovi klinično potrjenih bolnikov VBP smo izračunali skupno pojavnost VBP v Sloveniji, ki je 1 : 2.762, kar je primerljivo z ostalimi evropskimi državami. Zaključimo lahko, da smo z raziskavo uspešno določili ustrezne izključitvene vrednosti za posamezni izmerjeni presnovek in razmerja med njimi ter določili nabor VBP za slovenski program NBS. Pri oblikovanju strategije nacionalnega NBS smo ena redkih skupin, ki smo za potrditveno testiranje uporabili genetsko analizo NGS v kombinaciji z drugimi uveljavljenimi potrditvenimi testi pri VBP. Raziskava predstavlja tudi pomemben znanstveni prispevek na področju VBP, saj smo prepoznali in opredelili dve novi genetski spremembi ter za slovensko populacijo razvili inovativni sistem točkovanja za izbor preiskovancev z najvišjim tveganjem za VBP. Določili smo tudi pogostost VBP v Sloveniji, kar predstavlja prvo oceno pogostosti VBP v jugovzhodni Evropi.

Language:Slovenian
Keywords:biomedicina, farmacija, prirojene presnovne bolezni, zgodnja diagnostika, presejalni testi, novorojenčki, izbira kriterijev, ugotavljanje tveganj, Slovenija, ocene pogostosti, izbira metod, analize, doktorske disertacije
Work type:Dissertation
Typology:2.08 - Doctoral Dissertation
Organization:FFA - Faculty of Pharmacy
Place of publishing:Ljubljana
Publisher:[A. Šmon]
Year:2018
Number of pages:151 f.
PID:20.500.12556/RUL-137373 This link opens in a new window
UDC:616-008.9-056.7(043.3)
COBISS.SI-ID:293470464 This link opens in a new window
Publication date in RUL:15.06.2022
Views:614
Downloads:79
Metadata:XML RDF-CHPDL DC-XML DC-RDF
:
Copy citation
Share:Bookmark and Share

Secondary language

Language:English
Title:Definition criteria for expanded newborn screening for inborn errors of metabolism
Abstract:
Inborn errors of metabolism (IEM) are rare diseases; most have incidences lower than 1 : 10,000. Early detection of IEMs is crucial in making correct and timely clinical decisions that can prevent serious consequences of the diseases or even death. Contrary to many western European countries, most south-eastern European countries do not have an expanded newborn screening (NBS) program using tandem mass spectrometry and only screen for a few diseases using conventional methods like immunoassays and fluorimetry. The aim of the work was to prepare an optimal strategy for the organization of the expanded NBS for IEMs in Slovenia, based on selected criteria and scoring system that we designed for the purpose of this study. The criteria and the scoring system will be formulated and evaluated on the basis of the experimental results of our research and in this way we will determine the set of diseases that should be included in the expanded NBS program for the Slovenian population. We conducted a retrospective study where samples of dried blood spots on the filter paper of 10,048 subjects were analysed. In the initial NBS stage we identified each IEM with tandem mass spectrometry and confirmed it with confirmatory analyses. For confirmations we also used the next-generation sequencing method (NGS). This method allows the simultaneous analysis of all genes associated with IEMs and thus represents the method of choice for the confirmation of the diseases identified in the NBS. Due to unclear NBS results, 113 follow-ups were evaluated at an outpatient visit where also additional confirmatory analyses were performed. Confirmation tests included the analysis of dried blood spots on the filter paper, the analysis of organic acids in the urine, the analysis of amino acids in plasma, and in selected cases also the analysis of enzymatic activity in fibroblasts and acylcarnitine profiling in fibroblasts. On the basis of the results of 10,048 subjects analysis and comparisons with European countries, we determined the cut-off values for individual metabolites and the ratios between them, and developed a scoring system that allowed us to identify subjects with the highest risk of developing each of the selected IEMs. Based on this, we have proposed a set of IEMs that should be included in the expanded NBS program in Slovenia. With confirmatory tests four new patients with IEMs were detected, namely a patient with glutaric aciduria type 1, a patient with very long-chain-acyl-CoA-ehydrogenase deficiency and two patients with 3-methylcrotonyl-CoA-carboxylase deficiency. In all cases, the NGS approach identified causative genetic variants, such as those that have not yet been described in the literature. Previously undescribed genetic variants were the causative genetic variant c.205-7-8delCTinsGC in the ACADVL gene and the benign genetic variant c.149_151dupCCA (p.Thr50dup) in the MCCC1 gene. The estimated cumulative incidence of IEM in Slovenia, which was calculated from the patients detected in our study and from the number of clinically detected cases, is 1 : 2,762 and is comparable to other European countries. We can conclude that the study successfully determined the appropriate cut-off values for each measured metabolite and the ratios between them, and defined the set of IEMs for the Slovenian NBS program. We are one of the few groups that used the NGS genetic analysis for confirmatory testing in combination with other established confirmation tests for IEMs in the development of the national NBS strategy. The research also represents an important scientific contribution to the field of IEMs, as we identified and described two new genetic variants and for the Slovenian population developed an innovative scoring system for the selection of subjects with the highest risk for VBP. We have also determined the incidences of IEMs in Slovenia, which is the first estimate of the incidences of IEMs in South-Eastern Europe.


Projects

Funder:ARRS - Slovenian Research Agency
Project number:V3-1505
Name:Analiza in razvoj področja redkih bolezni v Sloveniji

Funder:ARRS - Slovenian Research Agency
Project number:P3-0343
Name:Etiologija, zgodnje odkrivanje in zdravljenje bolezni pri otrocih in mladostnikih

Similar documents

Similar works from RUL:
Similar works from other Slovenian collections:

Back