Halise Neslihan ÖNENLİ, Perçin PAZARCI, Ümit LÜLEYAP, Ayfer PAZARBAŞI, Davut ALPTEKİN, Halil KASAP, Gamze CÖMERTPAY, Ursula FROSTER

The analysis of the phenylalanine hydroxylase gene mutations by sequencing and ARMS techniques in Turkish patients

Amaç: Fenilketonüri, hepatik bir enzim olan fenilalanin hidroksilazın otozomal resesif bozukluğu sonucu ortaya çıkan bir durumdur. Bu çalışmada, Türk popülasyonunda en sık görülen fenilalanin hidroksilaz geni mutasyonlarını belirlemek amaçlandı. Gereç ve Yöntem: Öncelikle tüm ekson bölgeleri DHPLC metodu kullanılarak tarandı. Ardından mutasyon şüphesi taşıyan örnekler sekanslama ve ARMS yöntemleri kullanılarak analiz edildi. Bulgular: IVS10-11g->a splayzing mutasyonu 27 allelde (%58,7), R261Q mutasyonu 7 allelde (%15,2) ve E178G, P281L, R243X, R243Q, R408W, Y386C mutasyonları ise 2'şer allelde (%4,3) tespit edildi. Sonuç: R243X, R243Q, R261Q ve R408W mutasyonlarının oluştuğu 7 ve 12'inci ekson bölgelerinde arginin aminoasidinin değişimi Çukurova bölgesindeki fenilketonüri hastalarında fenilalanin hidroksilaz geni mutasyonlarında önemli bir yer tutmaktadır (%68,4). IVS10-11g->a ve P281L gibi tek baz mutasyonları bizim tarafımızdan dizayn edilmiş primerler ve ARMS tekniği kullanılarak daha ucuza, daha az teknik ekipmanla ve %100 doğrulukla tespit edilebilmektedir

Türk hastalarda fenilalanin hidroksilaz gen mutasyonlarının dizileme ve ARMS teknikleri ile incelenmesi

Purpose: Phenylketonuria is an autosomal recessive deficiency of the hepatic enzyme phenylalanine hydroxylase. With this study, detection of the most frequent phenylalanine hydroxylase gene mutations in Turkish population is aimed. Material and Methods: 23 unrelated phenylketonuria patients (46 alleles) who are living in Cukurova region, Turkey were investigated.First, all exons were screened by using DHPLC method then the direct sequencing technique and ARMS methods were used to analyze mutation suspected samples. Results: IVS10-11g->a splicing mutation in 27 alleles (58.7%), R261Q mutation in 7 alleles (15.2%) and E178G, P281L, R243X, R243Q, R408W, Y386C mutations, all in a frequency of 2/46 (4.3%) is found. Conclusion: The arginine amino acid, accounting for 68.4% of changes in exon sites 7 and 12, where R243X, R243Q, R261Q and R408W mutations occur, is thought to be important for amino acid changes in phenylalanine hydroxylase gene among phenylketonuria patients in Çukurova region. Single-base mutations like IVS10-11g->a and P281L could be detected with an accuracy of 100% by the use of specifically designed primers by authors according to ARMS technique and it is relatively cheaper and requesting less technical equipment

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