Renwick Pamela J, Trussler Jane, Ostad-Saffari Elham, Fassihi Hiva, Black Cheryl, Braude Peter, Ogilvie Caroline Mackie, Abbs Stephen
Genetics Centre, Guy's and St Thomas' NHS Foundation Trust, London SE1 9RT, UK.
Reprod Biomed Online. 2006 Jul;13(1):110-9. doi: 10.1016/s1472-6483(10)62024-x.
Preimplantation genetic haplotyping (PGH) proof-of-principle was demonstrated by multiple displacement amplification (MDA) of single buccal cells from a female donor and genotyping using 12 polymorphic markers within the dystrophin gene; the known paternal genotype enabled identification of the paternal haplotype in the MDA products despite 27% allele dropout. MDA amplified DNA from 49 single human blastomeres with 100% success. The MDA products were genotyped using a total of 57 polymorphic markers for chromosomes 1, 7, 13, 18, 21, X and Y; 72% of alleles amplified providing results at 90% of the loci tested. A PGH cycle was carried out for Duchenne muscular dystrophy. One embryo was biopsied: PGH showed a non-carrier female, which was transferred with no resulting pregnancy. A PGH cycle was carried out for cystic fibrosis. Seven embryos were biopsied and PGH allowed the exclusion of 2 affected embryos; a carrier and a non-carrier embryo were transferred resulting in an on-going twin pregnancy. PGH represents a paradigm shift in embryo diagnosis, as one panel of markers can be used for all carriers of the same monogenic disease, bypassing the need for development of mutation-specific tests, and widening the scope and availability of preimplantation genetic testing.
通过对一名女性供体的单个颊细胞进行多重置换扩增（MDA）并使用肌营养不良蛋白基因内的12个多态性标记进行基因分型，证明了植入前基因单倍型分析（PGH）的原理；尽管存在27%的等位基因缺失，但已知的父本基因型能够在MDA产物中鉴定出父本单倍型。MDA成功扩增了49个人类单细胞卵裂球的DNA。使用总共57个针对1、7、13、18、21、X和Y染色体的多态性标记对MDA产物进行基因分型；72%的等位基因被扩增，在所测试位点的90%获得结果。针对杜兴氏肌肉营养不良症进行了一个PGH周期。对一个胚胎进行活检：PGH显示为非携带者女性，该胚胎被移植但未成功妊娠。针对囊性纤维化进行了一个PGH周期。对7个胚胎进行活检，PGH排除了2个受影响的胚胎；移植了一个携带者胚胎和一个非携带者胚胎，导致正在进行的双胎妊娠。PGH代表了胚胎诊断的范式转变，因为一组标记可用于同一单基因疾病的所有携带者，无需开发突变特异性检测，从而扩大了植入前基因检测的范围和可及性。
