Yıl 2020, Cilt 3 , Sayı 3, Sayfalar 93 - 101 2020-11-05

Tanısı Zor Tek Gen Hastalıklarında Hedefe Yönelik Yeni Nesil Dizileme Panel Tasarımı: Primer İmmün Yetersizlik Örneği
Targeted Next Generation Sequencing Panel Design in Complex Monogenic Diseases: The Example of Primary Immunodeficiency

Sinem FIRTINA [1] , Özden HATIRNAZ NG [2] , Müge SAYİTOĞLU [3] , Yuk YİN NG [4]


Amaç: Yeni nesil dizileme teknolojileri bugün çok sayıda aday genin, genomun tüm kodlayan bölgelerinin hatta tüm genomun analizini tek seferde ve kısa süre içerisinde düşük maliyet ve yüksek hassasiyette, güvenilir bir şekilde mümkün kılmaktadır. Hedefe yönelik yeni nesil dizileme sistemleri genomda sadece belirli bölgenin dizilenmesine imkan veren, tüm genom dizilemelere göre uygulaması ve analizi daha kolay, hızlı ve yüksek güvenirlilikte bir yöntem olarak pek çok rutin genetik tanı uygulamalarında yerini bulmuştur. Gereç ve Yöntem: Çalışmamızda primer immün yetersizliklerin en yaygın grubu olan primer antikor yetersizlikleri (PAY) ve en ağır seyirli grubu ağır kombine immün yetersizlikler (AKİY) için hastalık ile ilişkili olduğu bilinen genleri kapsayan PZR temelli genetik tanı panelleri geliştirilmiş ve ortaya çıkan yüksek verinin yorumlanması için bir analiz akışı oluşturulmuştur. Bulgular: Tasarlanan paneller ile toplam 112 hasta (PAY:64, AKİY:48) dizilenmiş ve AKİY hastalarının %58’i ve PAY hastalarının %14,2’sinde hastalık ile ilişkili varyantlar tespit edilmiştir. Tüm varyantlar Sanger dizileme ile doğrulanarak oluşturulan moleküler tanı panellerinin ve analiz algoritmasının doğruluğu kontrol edilmiştir. Sonuç: Hedefe yönelik yeni nesil dizileme panellerinin hedeflenen bölgeye uygun olarak doğru yöntemle tasarlanması ve çıkan ham datanın doğru iş akışı ile analiz edilmesi panelin başarısını arttırmaktadır.

Objective: Next-generation sequencing technologies can generate an analysis of a large number of candidate genes, all the coding regions of a genome, or whole genomes with a high degree of accuracy and within a short amount of time. Targeted next generation sequencing systems have been found in many routine genetic diagnosis applications that allow the sequencing of only the candidate regions of the genome. Materials and Methods: In this study, we designed PCR-based targeted next generation sequencing (NGS) panels for severe combined immunodeficiency (SCID) and primary antibody deficiency (PAD) and created an algorithm for analysing high-throughput data. Results: We screened 112 patients (48 SCID and 64 PAD) and we detected genetic variations in 58% of the SCID and in 14.2% of the PAD patients. All variants were validated by Sanger sequencing to validate the accuracy of the NGS panel and analysis algorithm. Conclusion: Designing targeted next generation sequencing panels with an appropriate method, in accordance with the targeted region, and analysing the raw data with a suitable workflow, increases the success of the panel.

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Birincil Dil tr
Konular Temel Bilimler
Bölüm Research Article
Yazarlar

Orcid: 0000-0002-3370-8545
Yazar: Sinem FIRTINA
Kurum: İSTİNYE ÜNİVERSİTESİ
Ülke: Turkey


Orcid: 0000-0001-7728-6527
Yazar: Özden HATIRNAZ NG
Kurum: ACIBADEM ÜNİVERSİTESİ, TIP FAKÜLTESİ, TEMEL TIP BİLİMLERİ BÖLÜMÜ, TIBBİ BİYOLOJİ ANABİLİM DALI
Ülke: Turkey


Orcid: 0000-0002-8648-213X
Yazar: Müge SAYİTOĞLU
Kurum: İSTANBUL ÜNİVERSİTESİ, AZİZ SANCAR DENEYSEL TIP ARAŞTIRMA ENSTİTÜSÜ, GENETİK ANABİLİM DALI
Ülke: Turkey


Orcid: 0000-0001-9755-6045
Yazar: Yuk YİN NG (Sorumlu Yazar)
Kurum: İSTANBUL BİLGİ ÜNİVERSİTESİ, MÜHENDİSLİK VE DOĞA BİLİMLERİ FAKÜLTESİ, GENETİK VE BİYOMÜHENDİSLİK BÖLÜMÜ
Ülke: Turkey


Destekleyen Kurum istanbul üniversitesi BAP ve İstanbul Bilgi Üniversitesi araştırma fonu
Proje Numarası BAP: 52575 ve 20499, Bilgi Üniversitesi:NGYY-2018.01.0006
Teşekkür Çalışmaya dahil edilen hasta örneklerini sağlayan Doç. Dr. Selda Hançerli Torun, Prof. Dr. Elif Aydıner, Doç. Dr. Ayca Kiykim, Prof. Dr. Yildiz Camcioglu, Prof. Dr. Safa Baris ve Prof. Dr. Ahmet Ozen’e, Çalışmaya bilimsel katkıları için Prof. Dr. Ugur Ozbek’e teşekkürlerimizi sunarız.
Tarihler

Başvuru Tarihi : 7 Eylül 2020
Kabul Tarihi : 25 Eylül 2020
Yayımlanma Tarihi : 5 Kasım 2020

APA Fırtına, S , Hatırnaz Ng, Ö , Sayi̇toğlu, M , Yi̇n Ng, Y . (2020). Tanısı Zor Tek Gen Hastalıklarında Hedefe Yönelik Yeni Nesil Dizileme Panel Tasarımı: Primer İmmün Yetersizlik Örneği . Sağlık Bilimlerinde İleri Araştırmalar Dergisi , 3 (3) , 93-101 . Retrieved from https://dergipark.org.tr/tr/pub/sabiad/issue/57700/790469