公开(公告)号：US06924102B2

公开(公告)日：2005-08-02

申请号：US09938013

申请日：2001-08-24

申请人： Khue Vu Nguyen ,  Charles-Michel Wolff ,  Philippe Poindron

发明人： Khue Vu Nguyen ,  Charles-Michel Wolff ,  Philippe Poindron

IPC分类号： C12Q1/68 ,  C07H21/04

CPC分类号： C12Q1/6883 ,  C12Q2600/156 ,  C12Q2600/158

摘要： The present invention concerns the development of a quantitative method for the molecular diagnosis of autosomal recessive spinal muscular atrophy (SMA) by measuring the amount of cytosolic mRNA from human muscle cells. Both the procedure using radioactive material and the Enzyme-Linked Immunosorbent Assay (ELISA) nonradioactive method were developed using 32P-dCTP labeled and biotinylated nucleotide probes. The results obtained demonstrate that the measurement of mRNA could be used as a quantitative method for the molecular diagnosis of SMA. There was a perfect concordance of the results obtained between the procedure using radioactive material, the ELISA method and the single strand conformation polymorphism (SSCP) analysis regarding the negative and positive SMA samples. The methods developed in this study may be applicable to the diagnosis (detection of homozygous and heterozygous deletions in exons 7 and 8 of the SMN gene) and the control of mRNA concentrations in the future gene therapy of patients with SMA.

摘要翻译： 本发明涉及通过测量来自人肌肉细胞的胞质mRNA的量来开发用于常染色体隐性脊髓性肌萎缩（SMA）的分子诊断的定量方法。 使用放射性物质的方法和酶联免疫吸附测定（ELISA）非放射性方法都是使用32-p-dCTP标记和生物素化的核苷酸探针开发的。 获得的结果表明，mRNA的测量可以用作SMA的分子诊断的定量方法。 在使用放射性物质的方法，ELISA法和单链构象多态性（SSCP）分析之间获得的结果与阴性和阳性SMA样品完全一致。 本研究开发的方法可能适用于SMA患者未来基因治疗中的诊断（检测SMN基因外显子7和8中纯合和杂合缺失）和mRNA浓度的控制。