Sawada K, Doyu M, Tanaka F, Sobue G, Kato K
Taisho Laboratory of Functional Genomics, Nara Institute of Science and Technology, Nagoya, Japan.
Anal Biochem. 2000 Nov 1;286(1):59-66. doi: 10.1006/abio.2000.4786.
Triplet repeat disease is a group of hereditary neurodegenerative disorders caused by expansion of trinucleotide repeats such as CAG/CTG, CGG/CCG, and GAA/TTC. Direct detection of the expansion in the patient's genome shortcuts the tedious process needed for identification of disease genes by conventional approaches. Here we describe a method to detect triplet repeat expansion from the hybridization signal intensity. Using a digoxigenin-labeled (CTG)9 probe, the hybridization intensity and number of repeats showed a good linear correlation. The technique detected expansion in genomic DNA in all cases with moderate or large expansion. Even in the case of a small expansion, this method could detect the mutant fragment. The technique has advantages over related techniques because it is more sensitive and can be applied to cases where a small repeat expansion is involved.
三联体重复疾病是一组遗传性神经退行性疾病，由三核苷酸重复序列（如CAG/CTG、CGG/CCG和GAA/TTC）的扩增引起。直接检测患者基因组中的扩增可缩短通过传统方法鉴定疾病基因所需的繁琐过程。在此，我们描述一种从杂交信号强度检测三联体重复扩增的方法。使用地高辛标记的(CTG)9探针，杂交强度与重复次数显示出良好的线性相关性。该技术在所有中度或大量扩增的病例中均能检测到基因组DNA中的扩增。即使在小扩增的情况下，该方法也能检测到突变片段。该技术比相关技术具有优势，因为它更灵敏，可应用于涉及小重复扩增的病例。
