BRCA1 ve BRCA2 Mutasyonlarının Tespitine Yönelik Yeni Nesil Dizileme Temelli Kit Geliştirilmesi ve Rutinde Kullanılan Yöntemler ile Valide Edilmesi

Amaç: Meme kanseri, kadınlarda en yaygın görülen kanser türü olup, Göğüs Kanseri Duyarlılık gen (BRCA1 ve BRCA2) mutasyonlarının meme ve yumurtalık kanserlerinin önemli bir kısmından sorumlu olduğu bilinmektedir. Bu genlerden birinde mutasyon taşıyan bireylerde yaşam boyu meme, yumurtalık, pankreas ve diğer kanserlere yakalanma riski oldukça yükselmektedir. BRCA1/2 gen mutasyonlarına sahip olan kişilerin belirlenmesi, genetik danışma ile tarama sıklığının artırılması ve potansiyel olarak hayat kurtaran önleyici tedavi stratejilerinin uygulanabilmesi için büyük önem taşımaktadır. Bu çalışmada gerçek zamanlı polimeraz zincir reaksiyonu (RT-PZR) yöntemi kullanılarak BRCA1/2 genlerinin yeni nesil dizi (NGS) analizi kütüphanelerinin hazırlanması ve NGS analizlerine uygun biyoinformatik iş akışının belirlenmesi amaçlanmıştır.

Anahtar Kelimeler:

Meme Kanseri, BRCA1, BRCA2, Yeni Nesil Dizileme, NGS Kütüphane Oluşturması

Development of Next Generation Sequencing Based Kit for the Detection of BRCA1 and BRCA2 Mutations, and Validation With Routinely Used Methods

Aim: Breast cancer is the most common type of cancer in women, and it is well known that Breast Cancer Susceptibility gene (BRCA1 and BRCA2) mutations are responsible for a substantial portion of the breast and ovarian cancers. Individuals carrying a mutation in one of these genes have an increased substantially lifelong risk of breast, ovarian, pancreatic, and other types of cancers. Identifying individuals with BRCA1/2 gene mutations is vital for increasing screening of family members via genetic counselling, and using potentially life-saving preventive measures. The aim of this study is preparation of next generation sequencing (NGS) libraries of BRCA1/2 genes using real time polymerase chain reaction (RT-PCR), and determination of bioinformatic workflow suitable for NGS analysis.

Keywords:

Breast Cancer, BRCA1, BRCA2, Next Generation Sequencing, Library Preparation for NGS,

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