Department of Chemical and Biological Engineering, KOC University, Istanbul, Turkey.
Mol Cell Probes. 2010 Jun;24(3):138-41. doi: 10.1016/j.mcp.2009.12.001. Epub 2009 Dec 16.
Spinal muscular atrophy (SMA), the leading genetic cause of death in childhood, is an autosomal recessive neuromuscular disorder characterized by progressive muscle weakness, associated with deletions of the survival motor neuron 1 (SMN1) gene. Approximately 94% of SMA patients carry homologous deletions of SMN1 exon(s) 7 (and 8). Because of the high incidence and severity of the disease, precise detection and quantification of SMN1 and SMN2 gene copy numbers is essential for diagnosis and genetic counseling. We have developed a reliable single-tube tetra-primer PCR assay to simultaneously detect both the SMN1 and SMN2 exon 7 deletion using the advantage of C/T difference at nucleotide position of 840 in exon 7. The assay has been optimized and tested in 48 healthy controls, 20 known patients with SMA, 12 carriers (one SMN1 copy), and 8 amniotic fluids suspected of having SMA for whom we had determined the SMN1/SMN2 deletion by an additional PCR-RFLP method. We have observed complete concordance between methods. Our tetra-primer PCR assay is sensitive, low-cost, and easy to use method for simultaneous detection of both SMN1 and SMN2 deletion, which could be used even in "low-tech" laboratories.
脊髓性肌萎缩症（SMA）是儿童期主要的遗传致死原因，是一种常染色体隐性神经肌肉疾病，其特征是进行性肌肉无力，与生存运动神经元 1（SMN1）基因缺失有关。大约 94%的 SMA 患者携带 SMN1 外显子（s）7（和 8）的同源缺失。由于该疾病的高发病率和严重程度，精确检测和定量 SMN1 和 SMN2 基因拷贝数对于诊断和遗传咨询至关重要。我们开发了一种可靠的单管四引物 PCR 检测方法，利用外显子 7 核苷酸位置 840 的 C/T 差异，同时检测 SMN1 和 SMN2 外显子 7 的缺失。该检测方法已在 48 名健康对照、20 名已知 SMA 患者、12 名携带者（1 份 SMN1 拷贝）和 8 份疑似 SMA 的羊水进行了优化和测试，我们通过额外的 PCR-RFLP 方法确定了这些羊水的 SMN1/SMN2 缺失情况。我们观察到这两种方法之间完全一致。我们的四引物 PCR 检测方法是一种灵敏、低成本、易于使用的同时检测 SMN1 和 SMN2 缺失的方法，即使在“低技术”实验室也可以使用。
