Non-İnvaziv PGT Tekniği
Özet
Preimplantasyon genetik testi (PGT), tüp bebek (IVF) uygulamalarında elde edilen embriyolarda tek gen hastalıklarının (PGT-M, monogenik), translokasyon, delesyon/insersiyon veya inversiyon gibi kromozomal yapısal yeni düzenlenimlerin (PGT-SR, structural rearrengement) ve kromozomal sayı artış ya da azalışlarının (PGT-A, anöploidi) analizi için kullanılmaktadır. PGT-A uygulamaları ile hedeflenen klinik fayda, embriyolar arasında sayısal ve yapısal kromozomal anormallik taşımayanların transferi ile uterin implantasyon oranını arttırmak ve fetal düşük oranı azaltmaktır. Geleneksel olarak uygulanan PGT için invaziv yaklaşımla alınan hücre biyopsi örnekleri test edilmekte iken, embriyolardan in-vitro şartlarda içinde bulundukları kültür ortamına hücre dışı serbest DNA (cell free DNA, cfDNA) salındığının saptanması üzerine embriyoların blastokist evresindeki harcanmış kültür sıvısının (spent blastocyst media - SBM) invaziv olmayan bir yaklaşımla (noninvaziv PGT, niPGT) alternatif bir genetik materyal kaynağı olabileceği değerlendirilmiştir. NiPGT sonuçları, aynı embriyoya ait trofektoderm, iç hücre kütlesi (inner cell mass, ICM) veya tüm blastokist test sonuçlarıyla karşılaştırılmış ve zaman içerisinde test materyalinin uygunluğu (bilgi vericilik yönünde yeterli oranda bulunuşu) ve kromozomal ploidi sonuçları yönünden yüksek derecede anlamlı konkordans (uyum) gösteren veriler elde edilmiştir. NiPGT uygulamasında SBM içerisinde embriyoya ait olmayan cfDNA kontaminasyonunun engellemesi ve embriyoya ait cfDNA miktarını en üst düzeye çıkarılması için optimum sürede uzatılmış bir embriyo kültürü zamanına ihtiyaç vardır. Bu bölümde, SBM toplama protokolleri ve bilgi vericilik oranları, SBM örneklerinin referans örnekler ile karşılaştırılmasına dayanan uyum oranları, geleneksel PGT-A ve niPGT-A verilerinin karşılaştırılması, niPGT-A ile elde edilen klinik sonuçlar, transferde embriyo önceliklendirmesi için niPGT-A’nın potansiyel değeri ve SBM cfDNA kullanımındaki kısıtlılıklar ve zorluklar tartışılmıştır.
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