Abou-Sleiman P M, Apessos A, Harper J C, Serhal P, Delhanty J D A
UCL Centre for Preimplantation Genetic Diagnosis and the Assisted Conception Unit, Department of Obstetrics and Gynaecology, University College London, 86-96 Chenies Mews, London WC1E 6HX, UK.
Mol Hum Reprod. 2002 Mar;8(3):304-9. doi: 10.1093/molehr/8.3.304.
Crouzon syndrome is a dominantly inherited craniosynostosis syndrome which is caused by mutations in the fibroblast growth factor receptor 2 gene (FGFR2). However, a specific point mutation in the FGFR3 gene has also been shown to result in Crouzon syndrome associated with acanthosis nigricans. We report here the first method for preimplantation genetic diagnosis (PGD) of Crouzon syndrome based on multiplex PCR amplification followed by the direct detection of the causative mutation by single-stranded conformational polymorphism (SSCP) analysis. A highly polymorphic short tandem repeat (STR) locus was simultaneously analysed as a control against some forms of contamination. The mutation, carried by the female partner, was a de-novo substitution at codon 338 of the FGFR2 gene. The couple were found to be informative at the D21S11 STR locus. Two clinical PGD cycles were performed, resulting in the biopsy of 36 blastomeres, 25 of which showed amplification at the FGFR2 locus. All of the cells showed expected genotypes at the D21S11 locus with only one incidence of allele drop-out. A total of five embryos were transferred, two in the first cycle and three in the second, resulting in a singleton pregnancy.
克鲁宗综合征是一种常染色体显性遗传的颅缝早闭综合征,由成纤维细胞生长因子受体2基因(FGFR2)突变引起。然而,FGFR3基因中的特定点突变也已被证明会导致与黑棘皮病相关的克鲁宗综合征。我们在此报告了第一种用于克鲁宗综合征植入前基因诊断(PGD)的方法,该方法基于多重PCR扩增,随后通过单链构象多态性(SSCP)分析直接检测致病突变。同时分析了一个高度多态的短串联重复序列(STR)位点,作为针对某些形式污染的对照。女性伴侣携带的突变是FGFR2基因第338位密码子的新生替换。发现这对夫妇在D21S11 STR位点具有信息性。进行了两个临床PGD周期,共活检了36个卵裂球,其中25个在FGFR2位点显示扩增。所有细胞在D21S11位点均显示预期基因型,仅出现一次等位基因脱失。总共移植了5个胚胎,第一个周期移植2个,第二个周期移植3个,最终实现单胎妊娠。