Fiorentino F, Biricik A, Karadayi H, Berkil H, Karlikaya G, Sertyel S, Podini D, Baldi M, Magli M C, Gianaroli L, Kahraman S
EmbryoGen (Centre for Preimplantation Genetic Diagnosis), Rome, Italy.
Mol Hum Reprod. 2004 Jun;10(6):445-60. doi: 10.1093/molehr/gah055. Epub 2004 Mar 25.
Preimplantation HLA matching has recently emerged as a tool for couples desiring to conceive a potential donor progeny for transplantation in a sibling with a life-threatening disorder. In this paper we describe a strategy optimized for preimplantation genetic diagnosis (PGD) of haemoglobinopathies combined with HLA matching. This procedure involves a minisequencing-based genotyping of HLA regions A, B, C and DRB combined with mutation analysis of the gene regions involved by mutation. Analysis of at least eight polymorphic short tandem repeat (STR) markers scattered through the HLA complex has also been included to detect potential contamination and crossing-over occurrences between HLA genes. The above assay can also be used for preimplantation HLA matching as a primary indication. The strategy was clinically applied for HLA matching in 17 cycles (14 for beta-thalassaemia, one for Wiscott-Aldrich syndrome and two for leukaemia). A reliable HLA genotype was achieved in 255/266 (95.9%) of the blastomeres. In total, 22 (14.8%) embryos were obtained that were HLA-matched with the affected siblings, 14 (9.4%) of which were unaffected and transferred back to the patients. Four clinical pregnancies were obtained, three of which (one twin, two singletons) are ongoing and were confirmed as healthy and HLA-identical with the affected children. Minisequencing-based HLA typing combined with HLA STR haplotyping has been shown to be a reliable strategy for preimplantation HLA matching. The major advantage of this approach is that the validation of a single assay can be done once and then used for the majority of the patients, reducing notably time needed for preclinical set-up of each case.
植入前HLA配型最近已成为一种工具，供希望孕育潜在供体后代以移植给患有危及生命疾病的同胞的夫妇使用。在本文中，我们描述了一种针对血红蛋白病植入前基因诊断（PGD）并结合HLA配型进行优化的策略。该程序包括对HLA区域A、B、C和DRB进行基于微测序的基因分型，并结合对涉及突变的基因区域进行突变分析。还包括对散布在HLA复合体中的至少八个多态性短串联重复序列（STR）标记进行分析，以检测HLA基因之间的潜在污染和交叉事件。上述检测也可作为主要指征用于植入前HLA配型。该策略已在17个周期中临床应用于HLA配型（14例用于β地中海贫血，1例用于威斯科特-奥尔德里奇综合征，2例用于白血病）。在266个卵裂球中有255个（95.9%）获得了可靠的HLA基因型。总共获得了22个（14.8%）与患病同胞HLA匹配的胚胎，其中14个（9.4%）未受影响并移植回患者体内。获得了4例临床妊娠，其中3例（1例双胎，2例单胎）仍在继续，并被确认为健康且与患病儿童HLA相同。基于微测序的HLA分型结合HLA STR单倍型分型已被证明是一种可靠的植入前HLA配型策略。这种方法的主要优点是，单一检测的验证只需进行一次，然后可用于大多数患者，显著减少了每个病例临床前准备所需的时间。
