陈欣,孙宇,张凤山,潘胜发,王少波,张立,刁垠泽,周非非,赵衍斌,刘晓光,刘忠军

• 1:北京大学第三医院骨科

摘要(Abstract)：

[目的]建立目标基因测序技术,对后纵韧带骨化(ossification of the posterior longitudinal ligament,OPLL)患者的11个已知致病基因进行突变筛查,探讨OPLL与致病基因突变的关系。[方法]提取6例OPLL患者外周血全基因组DNA,利用Gen Cap目标基因捕获技术(北京迈基诺公司),设计骨形态发生蛋白-2(bone morphogenetic protein 2,BMP2)、骨形态发生蛋白-4(bone morphogenetic protein 4,BMP4)、骨形态发生蛋白-9(bone morphogenetic protein 9,BMP9)、Ⅺ型胶原蛋白α2(collagen type XI alpha 2,COL11A2)、Ⅵ型胶原蛋白α1(collagen type VI alpha1,COL6A1)、核苷酸焦磷酸酶(ectonucleotide pyrophosphatase/phosphodiesterase 1,ENPP1)、成纤维细胞生长因子2(fibroblast growth factor 2,FGF2)、成纤维细胞生长因子受体1(fibroblast growth factor receptor 1,FGFR1)、成纤维细胞生长因子受体2(fibroblast growth factor receptor 2,FGFR2)、转化生长因子-β3(transforming growth factor beta 3,TGFB3)和转化生长因子-β受体2(transforming growth factor beta receptor II,TGFBR2)基因外显子区域特异性捕获探针,与基因组DNA文库进行杂交,将目标基因组区域的DNA片段进行富集后,再利用illumina hiseq2000第二代测序仪进行测序,通过数据分析,确定突变位点,对选定的突变位点用Sanger测序法进行验证。[结果]设计合成的目标基因特异性捕获探针可有效地捕捉并富集基因组DNA的目标靶片段。目标区域平均测序深度为426.85~976.15,99.30%~100%目标区域覆盖度。在1例患者中发现COL6A1基因的1个错义突变,此位点检测结果与Sanger测序结果完全一致。[结论]目标基因测序技术成功发现了OPLL患者的致病基因突变。该方法快速而有效,对深入研究OPLL的分子病因学有重要意义。

关键词(KeyWords)： 后纵韧带骨化;致病基因;目标基因测序

Abstract:

Keywords:

基金项目(Foundation): 北京大学-清华大学生命科学联合中心交叉培育计划项目

作者(Author): 陈欣,孙宇,张凤山,潘胜发,王少波,张立,刁垠泽,周非非,赵衍斌,刘晓光,刘忠军

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