Primer monosemptomatik enürezis nokturnalı hastalarda akuaporin-2(AQP2) ve arginin vazopressin(AVP) genlerinin yeni nesil dizi analizi ile taranması
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Date
2021
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Düzce Üniversitesi
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Abstract
Giriş-Amaç: Enürezis nokturna çocukluk çağının en sık karşılaşılan üriner sistem problemlerinden biridir. Doğuştan ya da kazanılmış santral sinir sistemi defekti olmayan beş yaşın üzerindeki çocuklarda istemsiz olarak uykuda gece altını ıslatma enürezis nokturna olarak tanımlanır. Etiyolojisinde birçok faktörün rol aldığı bilinmekle birlikte fizyopatolojisinde üç önemli faktör rol oynar. Bunlar nokturnal poliüri, uyanma bozukluğu, gece düşük mesane kapasitesi ile aşırı detrusör aktivitesidir. Çocukluk çağında enürezis nokturna görülen ebeveynlerin çocuklarında hastalığın görülme riskinin arttığı bilinmektedir. Ancak literatürde primer monosemptomatik enürezis nokturnalı (PMEN) hastalarda akuaporin-2 ve arginin vazopressin genlerinin yeni nesil dizi analizi ile taramasının yapıldığı bir çalışma yoktur. Biz bu çalışma ile primer monosemptomatik enürezis nokturnalı hastalarda yeni nesil dizi analizi yöntemiyle etiyolojiden sorumlu genetik değişimi ortaya koymayı amaçladık. Gereç-Yöntem: Bu çalışma Düzce Üniversitesi Tıp Fakültesi Çocuk Sağlığı ve Hastalıkları Anabilim Dalı Çocuk Nefrolojisi Polikliniği'nde primer monosemptomatik enürezis nokturna olarak değerlendirilen 40 hasta ve 10 sağlıklı kontrol ile yapılmış kesitsel ve genetik bir çalışmadır. Olguların hedeflenen moleküler genetik tanısı için ilk olarak rutin amaçlı istenilen tam kan örneklerinden geriye kalan atık kanlardan DNA izolasyon işlemi yapıldı. İzole edilen DNA örneklerinden hedeflenen gen bölgeleri, bu bölgelere spesifik primerler kullanılarak çoğaltılıp saflaştırma işlemi uygulandıktan sonra hedefe yönelik yeni nesil dizi analizi yöntemiyle bu bölgelerdeki mutasyonlar saptandı. Bulgular: Primer monosemptomatik enürezis nokturna (PMEN) tanısı alan 40 hastanın 23'ü kız (%57,5), 17'si ise erkek (%42,5) idi. Kız/erkek oranı 1,35/1 olarak hesaplandı. Hasta grubunda 23 (%57,5) çocukta ailede enürezis öyküsü belirlendi. Hasta grubunda 30 (%75) hastanın ikinci derece akrabasında enürezis öyküsü tespit edildi. Çalışmamıza dahil olan deneklerde AQP2 geninde ekzon 1'de c.360+3G>A heterozigot, c.360+3G>A homozigot, V13=c.39G>A heterozigot varyasyonları, ekzon 2'de S167=c.501T>C heterozigot, S167=c.501T>C homozigot varyasyonları, ekzon 4'te c.816+20C>A heterozigot, c.816+20C>A homozigot varyasyonları saptanmıştır. AVP geninde ise sadece ekzon 2'de c.322+2T>G homozigot ve R51H c.152G>A heterozigot varyasyonları saptanmıştır. Sonuç: AQP2 ekzon 1'deki c.360+3G>A homozigot varyasyonu ve AVP geni ekzon 2'deki c.322+2T>G homozigot varyasyonuna sadece PMEN'lı çocuklarda rastlanmış, kontrol grubundaki çocuklarda rastlanılmamış olması nedeniyle tespit edilen bu değişikliklerin PMEN için belirteç olarak kullanılabileceğini düşünüyoruz. Anahtar Kelimeler: Enürezis nokturna, yeni nesil dizileme, akuaporin-2 (AQP2), arginin vazopresssin (AVP), çocuk
Introduction-Aim: Enuresis nocturna is one of the most common urinary system disorders of childhood. In children over five years of age who have no congenital or acquired central nervous system defects, nighttime wetting is defined as nocturnal enuresis nocturna. Although it is known that many factors play a role in its etiology, three important factors play a role in its physiopathology. These are nocturnal polyuria, awakening disorder, excessive detrusor activity with low bladder capacity at night. It is known that the risk of the enuresis is commonly seen in the children of the parents who have enuresis nocturna in their children. However, in the literature, there is no study in which aquaporin-2 (AQP2) and arginine vasopressin (AVP) genes were screened by a new generation sequence analysis in primary monosymptomatic enuresis nocturna patients. In this study, we aimed to reveal the genetic changes responsible for etiology by a new generation of sequence analysis method in patients with primary monosymptomatic enuresis. Material-Method: This study was a cross-sectional and genetic study conducted with 40 patients who were evaluated as primary monosymptomatic enuresis nocturna and 10 healthy controls in Düzce University Faculty of Medicine, Department of Pediatrics, Pediatric Nephrology Outpatient Clinic. For the targeted molecular genetic diagnosis of the cases, DNA isolation was firstly performed from the wasted blood remaining from the routine whole blood samples. The targeted gene regions from the isolated DNA samples were amplified and purified using primers specific to these regions and mutations in these regions were determined by targeted next generation sequencing. Results: Of the 40 patients diagnosed with primary monosymptomatic enuresis nocturna (PMEN), 23 were female (57.5%) and 17 were male (42.5%). The female / male ratio was calculated as 1.35 / 1. In the patient group, 23 (57.5%) children had a family history of enuresis. In the patient group, 30 (75%) patients' second degree relatives had a history of enuresis. In the subjects included in our study, in the AQP2 gene c.360 + 3G> A heterozygous, c.360 + 3G> A homozygous, V13=c.39G>A heterozygous variations in exon 1, S167 = c.501T> C heterozygous, S167 = c.501T> C homozygous variations in exon 2 , c.816 + 20C> A heterozygous, c.816 + 20C> A homozygous variations were detected in exon 4. In the AVP gene, c.322 + 2T> G homozygous and R51H c.152G> A heterozygous variations were detected only in exon 2. Conclusion: The c.360 + 3G> A homozygous variation in AQP2 exon 1 and the c.322 + 2T> G homozygous variation in the AVP gene exon 2 were found only in children with PMEN, and these changes were not found in children in the control group. Therefore, we thought that these detected changes might be used as markers for PMEN. Keywords: Enuresis nocturna, next generation sequencing analysis, aquaporin-2 (AQP2), arginine vasopressin (AVP), child
Introduction-Aim: Enuresis nocturna is one of the most common urinary system disorders of childhood. In children over five years of age who have no congenital or acquired central nervous system defects, nighttime wetting is defined as nocturnal enuresis nocturna. Although it is known that many factors play a role in its etiology, three important factors play a role in its physiopathology. These are nocturnal polyuria, awakening disorder, excessive detrusor activity with low bladder capacity at night. It is known that the risk of the enuresis is commonly seen in the children of the parents who have enuresis nocturna in their children. However, in the literature, there is no study in which aquaporin-2 (AQP2) and arginine vasopressin (AVP) genes were screened by a new generation sequence analysis in primary monosymptomatic enuresis nocturna patients. In this study, we aimed to reveal the genetic changes responsible for etiology by a new generation of sequence analysis method in patients with primary monosymptomatic enuresis. Material-Method: This study was a cross-sectional and genetic study conducted with 40 patients who were evaluated as primary monosymptomatic enuresis nocturna and 10 healthy controls in Düzce University Faculty of Medicine, Department of Pediatrics, Pediatric Nephrology Outpatient Clinic. For the targeted molecular genetic diagnosis of the cases, DNA isolation was firstly performed from the wasted blood remaining from the routine whole blood samples. The targeted gene regions from the isolated DNA samples were amplified and purified using primers specific to these regions and mutations in these regions were determined by targeted next generation sequencing. Results: Of the 40 patients diagnosed with primary monosymptomatic enuresis nocturna (PMEN), 23 were female (57.5%) and 17 were male (42.5%). The female / male ratio was calculated as 1.35 / 1. In the patient group, 23 (57.5%) children had a family history of enuresis. In the patient group, 30 (75%) patients' second degree relatives had a history of enuresis. In the subjects included in our study, in the AQP2 gene c.360 + 3G> A heterozygous, c.360 + 3G> A homozygous, V13=c.39G>A heterozygous variations in exon 1, S167 = c.501T> C heterozygous, S167 = c.501T> C homozygous variations in exon 2 , c.816 + 20C> A heterozygous, c.816 + 20C> A homozygous variations were detected in exon 4. In the AVP gene, c.322 + 2T> G homozygous and R51H c.152G> A heterozygous variations were detected only in exon 2. Conclusion: The c.360 + 3G> A homozygous variation in AQP2 exon 1 and the c.322 + 2T> G homozygous variation in the AVP gene exon 2 were found only in children with PMEN, and these changes were not found in children in the control group. Therefore, we thought that these detected changes might be used as markers for PMEN. Keywords: Enuresis nocturna, next generation sequencing analysis, aquaporin-2 (AQP2), arginine vasopressin (AVP), child
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Keywords
Enürezis nokturna, yeni nesil dizileme, akuaporin-2 (AQP2), arginin vazopresssin (AVP), çocuk, Enuresis nocturna, next generation sequencing analysis, aquaporin-2 (AQP2), arginine vasopressin (AVP), child, Çocuk Sağlığı ve Hastalıkları, Child Health and Diseases, Akuaporinler, Aquaporins, Arjinin, Arginine, Dizi analizi, Sequence analysis, Enürezis, Enuresis, Genler, Genes, Nokturnal enürezis, Nocturnal enuresis, Vazopressinler, Vasopressins
