Sermon K, De Vos A, Van de Velde H, Seneca S, Lissens W, Joris H, Vandervorst M, Van Steirteghem A, Liebaers I
Centre for Medical Genetics, University Hospital and the Medical School of the Dutch-speaking Brussels Free University, Belgium.
Mol Hum Reprod. 1998 Aug;4(8):791-6. doi: 10.1093/molehr/4.8.791.
Myotonic dystrophy (DM), or Steinert's disease, is an autosomal dominant disease characterized by myotonia, muscular weakness and atrophy, as well as lens opacities, cardiomyopathy and mild endocrine changes. The gene for DM located on 19q contains a triplet repeat at the 3' end of the gene. In DM patients, this repeat is found to be expanded. We have previously described a preimplantation genetic diagnosis (PGD) for DM using polymerase chain reaction (PCR) followed by conventional analysis on ethidium bromide-stained gels. The major drawback of this system was that allelic dropout occurred in >20% of the cells, leading to the loss of healthy embryos for transfer. To resolve this problem, we developed a PGD for DM using fluorescent PCR followed by fragment analysis on an automated DNA sequencer and made a comparison between the conventional PCR described earlier and fluorescent PCR, which turned out to be superior in accuracy and efficiency. Three PGD cycles were performed using fluorescent PCR and are described here.
强直性肌营养不良（DM），即斯坦纳特病，是一种常染色体显性疾病，其特征为肌强直、肌肉无力和萎缩，以及晶状体混浊、心肌病和轻度内分泌变化。位于19号染色体长臂（19q）上的DM基因在基因的3'末端含有一个三联体重复序列。在DM患者中，发现这个重复序列有所扩增。我们之前描述过一种针对DM的植入前基因诊断（PGD）方法，该方法先采用聚合酶链反应（PCR），然后在溴化乙锭染色的凝胶上进行常规分析。该系统的主要缺点是等位基因脱落在超过20%的细胞中发生，导致可供移植的健康胚胎丢失。为了解决这个问题，我们开发了一种针对DM的PGD方法，采用荧光PCR，随后在自动DNA测序仪上进行片段分析，并对之前描述的常规PCR和荧光PCR进行了比较，结果表明荧光PCR在准确性和效率方面更具优势。这里描述了使用荧光PCR进行的三个PGD周期。
