1. Detection of duchene muscular dystrophy carriers with quantitative fluorescent polymerase chain reaction
- Author
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Seniha Hacihanefioglu, Aysegul Kuskucu, Piraye Oflazer, Nur Buyru, Kuşkucu, Ayşegül, Buyru, Nur, Oflazer, Piraye, Hacıhanefioğlu, Seniha, and Yeditepe Üniversitesi
- Subjects
Mikrobiyoloji ,Duchenne muscular dystrophy ,Tıbbi Laboratuar Teknolojisi ,Nörolojik Bilimler ,Yoğun Bakım ,law.invention ,Exon ,Diş Hekimliği ,Transplantasyon ,law ,Multiplex ,Anatomi ve Morfoloji ,Enfeksiyon Hastalıkları ,Patoloji ,Sağlık Bilimleri ve Hizmetleri ,Muscular dystrophy ,Polymerase chain reaction ,Cerrahi ,Dermatoloji ,Alerji ,Hücre Biyolojisi ,Odyoloji ve Konuşma-Dil Patolojisi ,Kulak ,General Medicine ,Tıbbi İnformatik ,Amplicon ,Toksikoloji ,Kadın Hastalıkları ve Doğum ,Onkoloji ,Periferik Damar Hastalıkları ,Psikiyatri ,Tıp ,Tropik Tıp ,Gastroenteroloji ve Hepatoloji ,Romatoloji ,Biyoloji ,Temel Sağlık Hizmetleri ,Biyoteknoloji ve Uygulamalı Mikrobiyoloji ,Nükleer Tıp ,Kalp ve Kalp Damar Sistemi ,Burun ,Endokrinoloji ve Metabolizma ,Anestezi ,Biology ,Biyofizik ,Radyoloji ,Klinik Nöroloji ,Tıbbi Görüntüleme ,Üroloji ve Nefroloji ,Solunum Sistemi ,Acil Tıp ,Androloji ,Multiplex polymerase chain reaction ,medicine ,Spor Bilimleri ,Halk ve Çevre Sağlığı ,Multiplex ligation-dependent probe amplification ,Genel ve Dahili Tıp ,Geriatri ve Gerontoloji ,Fizyoloji ,Göz Hastalıkları ,İmmünoloji ,Farmakoloji ve Eczacılık ,Boğaz ,Tıbbi Araştırmalar Deneysel ,medicine.disease ,Tıbbi Etik ,Molecular biology ,Adli Tıp ,Pediatri ,Ortopedi ,Hematoloji ,Rehabilitasyon ,Tamamlayıcı ve Entegre Tıp ,Biyokimya ve Moleküler Biyoloji - Abstract
Aim: Duchenne Muscular Dystrophy (DMD) is an X-linked, progressive, lethal neuromuscular disorder affecting 1/3500 live-born males. Mutations occur in the dystrophin gene, which is located at Xp21.2. Partial gene deletions occur in two “hot-spot” regions, and can be responsible for up to 60-65% DMD cases, while 5-10% of the cases are caused from clustered gene duplications. Mutations can be inherited from female carriers (2/3) or be de-novo mutations (1/3). Deletions can be easily detected in affected males via multiplex PCR or MLPA. On the contrary, determining the status of female carriers is difficult. The aim of this study is to optimize the gene-dosage method using quantitative fluorescent PCR. Material and Methods: Fluorescently labeled primers are used for amplification and automated detection of amplicons and are designed in multiplex format. The primers contain eighteen exons located within “hot-spot” regions. A promoter region and STR markers are also included in the test as internal controls and for linkage analysis. This is followed by a PCR automated genetic analyzer for the detection of PCR products. This study includes twenty-four families, each with a previously diagnosed member. Results: Results showed the same correlation as was previously reported in nineteen patients, whereas three patients had an extra exon deletion and one patient had one less exon deletion than previously reported. In nineteen families, the mothers were carriers, and in five families, the mothers were not carriers. Conclusion: As a conclusion for carrier screening in DMD patients, quantitative fluorescent PCR is a fast, reproducible and robust method can be used for detection of deletions.
- Published
- 2019
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