20.500.12556/DKUM-88275 Detection of aneuploidy using multiplex ligation-dependent probe amplification in fetal tissues from aborted pregnancies Določanje anevploidij z metodo pomnoževanja od ligacije odvisnih prob v fetalnih tkivih splavkov Purpose: About 10-15% of all pregnancies terminate as spontaneous miscarriages. In the first trimester, 50% of spontaneous miscarriages are the result of chromosomal aberrations, mostly chromosomal aneuploidies. Cytogenetic analyses are used to confirm aneuploidy in failed pregnancies. Culture failure or poor quality chromosomes are often problems in those cases. In such situations, methods that are independent of tissue culture areused, and we employed multiplex ligation-dependent probe amplification (MLPA). We determined if MLPA is an appropriate and compatible method compared with classical cytogenetic analyses on fetal tissues. Methods: All fetal samples received from spontaneous abortions were cultured, karyotyped (if possible) and genomic DNA extracted. MLPA analyses were undertaken using subtelomeric probe kits. Additionally, comparative genomic hybridization (CGH) was used to confirm aneuploidy detected by MLPA in cases of failed culture growth. Results: MLPA analyses confirmed an unbalanced chromosome abnormality identified by cytogenetic analyses in all cases in which tissue culture was successful, and provided data in cases of failed culture growth. Several common numeric chromosome aberrations were detected, as well as rare trisomies and other unbalanced chromosome rearrangements. Conclusions: MLPA analyses can provide information about the karyotype of a DNA sample if cytogenetic analyses are not possible because of a lack of viable cells or if only a small amount of genomic DNA is available. These data indicate that MLPA may also be a very useful method for early prenatal aneuploidy screening. Namen: Spontani splavi se pojavljajo v približno 10-15% prepoznavnih nosečnosti. V prvem trimesečju je približno 50% splavov posledica kromosomskih napak, v večini primerov so to kromosomske anevploidije. Klasična metoda določanja anevploidij je citogenetska analiza. Citogenetska analiza zgodnjih spontanih splavov je težavna zaradi pogoste odsotnosti celične rasti ali slabe kvalitete kromosomov. V teh primerih se poslužujemo drugih metod, neodvisnih od rasti celične kulture. V študiji smo uporabili metodo pomnoževanja od ligacije odvisnih prob (MLPA). V primerjavi s klasično citogenetsko analizo smo na vzorcu embrionalnih tkiv potrdili ustreznost in kompatibilnost metode. Metode: Vsi vzorci embrionalnih tkiv po spontanih splavih so bili kultivirani, kariotipizirani, prav tako je bila izolirana genomska DNA. Za MLPA analizo smo uporabili komercialne komplete s subtelomerno specifičnimi DNA sondami. V primeru odsotnosti celične rasti so bile anevploidije ugotovljene z MLPA analizo, potrjene s primerjalno genomsko hibridizacijo (PGH). Rezultati: MLPA analiza je potrdila neuravnotežene kromosomske nepravilnosti, ugotovljene s citogenetsko analizo pri vseh vzorcih, kjer je bila uspešna celična rast, in hkrati omogočila analizo v primerih, kjer celična rast ni bila uspešna. Ugotovljene so bile mnoge številčne kromosomske spremembe, redke trisomije in druge neuravnotežene kromosomske preureditve. Zaključek: MLPA analiza omogoča pridobitev informacij o številu kromosomov v primerih, ko citogenetska analiza ni možna zaradi odsotnosti celične rasti ali slabe kvalitete kromosomov. Iz dobljenih rezultatov ugotavljamo, da je MLPA potencialno tudi zelo uporabna metoda za hitro in kvalitetno prenatalno diagnostiko. pomnoževanje od ligacije odvisnih prob številčne kromosomske preureditve kariotip multiplex ligation–dependent probe amplification numeric chromosome aberration karyotype true true false Angleški jezik Slovenski jezik Znanstveno delo 2024-04-12 14:51:01 2024-04-12 14:51:03 2026-03-19 13:54:56 0000-00-00 00:00:00 2011 0 0 str. 51-60 št. 2 Letn. 4 2011 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2011-01-01 618.39-021.3:575 4160831 1855-5640 http://www.actamedbio.mf.uni-mb.si/pdf/06%207id_amb_%2042_11.pdf 1 543f56fb-f8cb-11ee-a0d6-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=178485 06.pdf 06.pdf 1 13EE1174542C0BA259339ED0AE6D7093 63e4eadfb13d1c5f6ac76063764cd6694f23f275fcc05344995efba63af83238 9caa0642-f8cb-11ee-a0d6-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=178486 Medicinska fakulteta Acta medico-biotechnica 0 0 0