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標題: | 次世代定序在膠原蛋白病變的臨床應用 Clinical Application of Next Generation Sequencing in the Diagnosis of Collagenopathies |
作者: | Ming-Cheng Kuo 郭明正 |
指導教授: | 簡穎秀(Yin-Hsiu Chien 簡穎秀) |
關鍵字: | 膠原蛋白,膠原蛋白病變,異質性遺傳疾病,次世代定序,目標區間定序套組,外顯子定序套組, collagen,collagenopathy,heterogeneous genetic disorder,next generation sequencing,NGS-based targeted gene panel,Exome panel, |
出版年 : | 2019 |
學位: | 碩士 |
摘要: | 背景
膠原蛋白是人體結締組織中含量最豐富的蛋白質,由於膠原蛋白病變是異質性遺傳疾病(heterogeneous genetic disorder),許多不同基因變異可能造成相同類似的症狀。因此,在診斷這類疾病往往需要檢測不同基因,才能找到致病原因。傳統桑格定序(Sanger sequencing)對於處理多個基因或外顯子顯得耗費時間和成本,因此,次世代定序(Next Generation Sequencing;NGS)則能同時提供多個基因進行平行定序,具有縮短時間和降低成本的優勢。 目的 我們希望藉由次世代定序目標區間定序套組(NGS-based targeted gene panel),對於疑似膠原蛋白病變的患者,提供精確快速有效率的臨床分子診斷,並進一步去了解膠原蛋白病變的基因型和表現型相關性。 方法 本次研究收錄從2016年1月到2018年12月這段期間,疑似膠原蛋白病變的30位患者,其中有26位患者使用次世代定序目標區間定序套組,4位進行外顯子定序套組(Exome panel)檢測。8例臨床診斷為Elhers-Dalnos syndrome (EDS),22例臨床診斷為骨骼發育不良Skeletal dysplasia。 結果 19位患者有確定的分子診斷,診斷率為63% (n=19/30)。8位在臨床診斷為EDS患者,有4位找到致病基因,分別是1位PRDM5(Brittle cornea syndrome 2),2 位COL5A1(Classical EDS,cEDS)和1位COL3A1(Vascular EDS,vEDS)變異,診斷率為50%(n= 4/8)。22位骨骼發育不良患者,有4位是COL1A1和1位是COL1A2(Osteogenesis imperfect,OI)變異,4位是COL2A1 (COL2A1-related collagenopathy),其他基因型包括SERPINF1(OI type VI,n = 2),WNT1(OI type XV,n = 1),ITIM5(OI type V,n = 1),TRPV4(Spondylometaphyseal dysplasia,Kozlowski型,n = 1)和B3GALT6(Spondyloepimetaphyseal dysplasia with joint laxity,n = 1),診斷率為68%(n=15/22)。 結論 本次研究使用次世代定序骨骼關節疾病套組(OI、EDS、Osteopetrosis、Marfanoid、others)相關基因共56個,相較其他相關基因研究套組有較佳的分子診斷率。因此,可以做為臨床有效的診斷工具和成為診斷膠原蛋白病變的標準方法。 Background Collagen is the most abundant protein found in the connective tissue of the human body. Collagenopathy is a heterogeneous genetic disease, and many different genetic variations may cause similar symptoms. Sanger sequencing is too time-consuming and costly for processing multiple genes or exons. Next-generation sequencing (NGS) enables massive parallel sequencing of multiple genes at the same time, and it also has the advantage of reducing time and cost. Aim We try to apply the NGS-based targeted gene panel and offer accurate, fast and effective clinical molecular diagnosis for patients who were suspected with collagenopathies, and to further understand the correlation between genotypes and phenotypes. Methods The study was included thirty patients suspected of having collagenopathies from January 2016 to December 2018, and twenty-six of them used the NGS-based targeted gene panel. Four cases were tested by Exome panel. Eight of them were diagnosed as Elhers-Dalnos syndrome (EDS), and the other were distinguished as skeletal dysplasia. Results There were nineteen patients with molecular diagnosis, and the diagnostic rate was 63% (n=19/30). There were four patients with genetic variations in the clinical diagnosis of EDS, including PRDM5 (Brittle cornea syndrome 2, n=1), COL5A1 (Classical EDS,cEDS, n=2) and COL3A1 (Vascular EDS,vEDS, n=1) with the diagnostic yield of 50% (n=4/8). Twenty-two patients were diagnosed with skeletal dysplasia, four cases were COL1A1 and one case was COL1A2 (Osteogenesis imperfect, OI) variation, four cases were COL2A1(COL2A1-related collagenopathy), and other genotypes were included SERPINF1 (OI type VI, n = 2), WNT1 (OI type XV, n = 1), ITIM5 (OI type V, n = 1), TRPV4 (Spondylometaphyseal dysplasia, Kozlowski type, n = 1) and B3GALT6 (Spondyloepimetaphyseal dysplasia with joint laxity, n = 1). The diagnosis rate was 68% (n=15/22). Conclusions This study was utilized to fifty-six genes related to skeletal joint disease (OI、EDS、Osteopetrosis、Marfanoid、others) in the NGS-based targeted gene panel, which has a better molecular diagnostic rate than other related studies so that it can be used as a clinically effective diagnostic tool and standards for diagnosis of collagenopathies. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/7214 |
DOI: | 10.6342/NTU201902431 |
全文授權: | 同意授權(全球公開) |
顯示於系所單位: | 分子醫學研究所 |
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