Eftedal I, Schwartz M, Bendtsen H, Andersen A N, Ziebe S
The Fertility Clinic, Rigshospitalet, University Hospital Copenhagen, Blegdamsvej 9, DK-2100 Copenhagen, Denmark.
Mol Hum Reprod. 2001 Mar;7(3):307-12. doi: 10.1093/molehr/7.3.307.
This study is part of a strategy aimed at using fluorescent polymerase chain reaction (PCR) on informative genetic microsatellite markers as a diagnostic tool in preimplantation genetic diagnosis (PGD) of severe monogenic disease. Two couples, both of whom had previously had children who were compound heterozygote for severe cystic fibrosis mutations, were offered PGD using fluorescent PCR of the highly polymorphic cystic fibrosis transmembrane conductance regulator (CFTR) intragenic microsatellite marker IVS17bTA. Cleavage-stage embryo biopsy followed by PCR resulted in transfer of one unaffected carrier embryo for each couple. This approach eliminates the need for single cell multiplex PCR strategies to detect CF compound heterozygotes. It also provides a control of chromosome 7 ploidy in the blastomeres and a selection against allele dropout by positive detection of each CFTR copy of all genotypes in preimplantation embryos from genetically informative families.
本研究是一项策略的一部分，该策略旨在对信息丰富的基因微卫星标记物进行荧光聚合酶链反应（PCR），作为严重单基因疾病植入前基因诊断（PGD）的诊断工具。两对夫妇，他们之前都有孩子是严重囊性纤维化突变的复合杂合子，被提供使用高度多态性囊性纤维化跨膜传导调节因子（CFTR）基因内微卫星标记IVS17bTA的荧光PCR进行PGD。卵裂期胚胎活检后进行PCR，结果是每对夫妇都移植了一个未受影响的携带者胚胎。这种方法消除了检测CF复合杂合子所需的单细胞多重PCR策略。它还能对卵裂球中的7号染色体倍性进行控制，并通过在有遗传信息的家庭的植入前胚胎中对所有基因型的每个CFTR拷贝进行阳性检测来选择避免等位基因脱失。
