公开(公告)号：US6136546A

公开(公告)日：2000-10-24

申请号：US57740

申请日：1998-04-09

申请人： Kevin P. Campbell ,  Valerie Allamand ,  Yoshihide Sunada ,  Volker Straub ,  Mustafa Salih

发明人： Kevin P. Campbell ,  Valerie Allamand ,  Yoshihide Sunada ,  Volker Straub ,  Mustafa Salih

IPC分类号： G01N33/68 ,  G01N33/53 ,  G01N1/30 ,  G01N33/567

CPC分类号： G01N33/6887 ,  G01N2800/2878

摘要： Disclosed are compositions and methods for aiding in the diagnosis of congenital muscular dystrophy associated with in-frame deletion in the laminin-2 .alpha.2 polypeptide chain in an individual. In a preferred diagnostic method embodiment, an experimental muscle tissue sample is provided from the individual and treated if necessary to render components available for antibody binding. The components of the sample are then separated on the basis of molecular weight. The separated protein components are then transferred to a solid support while maintaining the relative positions established in separation step. The transferred components are then stained with an affinity reagent which is known to bind to a C-terminal domain of the laminin-2 .alpha.2 polypeptide chain. Individual afflicted with congenital muscular dystrophy associated with in-frame deletion in the laminin-2 .alpha.2 polypeptide chain on the basis of positive staining in combination with reduced molecular weight of the laminin-2 .alpha.2 polypeptide chain relative to the wild-type laminin-2 .alpha.2 polypeptide chain. A preferred composition is a nucleic acid probe for the detection of merosin deletion-type congenital muscular dystrophy. The preferred nucleic acid probe is characterized by the ability to bind specifically to a mutant merosin nucleic acid sequence, the mutant merosin nucleic acid sequence comprising a T to C substitution at position 3973 +2 of the consensus donor splice site of exon 25.

摘要翻译： 公开了用于帮助诊断个体中层粘连蛋白-2α2多肽链中与框内缺失相关的先天性肌营养不良的组合物和方法。 在优选的诊断方法实施方案中，从个体提供实验肌肉组织样品，并且如果需要进行处理以使组分可用于抗体结合。 然后基于分子量分离样品的组分。 然后将分离的蛋白质组分转移到固体支持物中，同时保持在分离步骤中建立的相对位置。 转移的组分然后用已知结合层粘连蛋白-2α2多肽链的C末端结构域的亲和试剂染色。 基于阳性染色结合层粘连蛋白-2α2多肽链相对于野生型层粘连蛋白的层粘连蛋白-2α2多肽链的分子量降低而与层粘连蛋白-2α2多肽链中的框内缺失相关的先天性肌营养不良患者， 2α2多肽链。 优选的组合物是用于检测梅洛辛缺失型先天性肌营养不良症的核酸探针。 优选的核酸探针的特征在于特异性结合突变型子糖蛋白核酸序列的能力，所述突变蛋清核酸序列包含外显子25的共有供体剪接位点的第3973±2位的T至C取代。