Zeschnigk M, Lich C, Buiting K, Doerfler W, Horsthemke B
Institut für Genetik, Universität zu Köln, Deutschland.
Eur J Hum Genet. 1997 Mar-Apr;5(2):94-8.
The analysis of allelic methylation differences in 15q11-q13 has been established as a valid test for the Angelman and Prader-Willi syndromes. Current tests use methylation-sensitive restriction enzymes and Southern blot analysis. Here we describe a single-tube PCR test. It is based on sodium bisulfite treatment of DNA, which converts unmethylated, but not methylated cytosine residues to uracil, and PCR primers specific for the maternal and the paternal allele. The method was validated in a blinded retrospective study on 87 DNA samples from normal controls and patients. Prospective studies by independent laboratories will be needed before this assay can replace Southern blot analysis in routine diagnostic procedures.
15q11 - q13等位基因甲基化差异分析已被确立为检测安吉尔曼综合征和普拉德 - 威利综合征的有效方法。目前的检测方法使用甲基化敏感限制酶和Southern印迹分析。在此，我们描述一种单管PCR检测方法。它基于对DNA进行亚硫酸氢钠处理，该处理可将未甲基化而非甲基化的胞嘧啶残基转化为尿嘧啶，以及针对母本和父本等位基因的特异性PCR引物。该方法在一项对87份来自正常对照和患者的DNA样本的盲法回顾性研究中得到验证。在该检测方法能够在常规诊断程序中取代Southern印迹分析之前，还需要独立实验室进行前瞻性研究。
