Jiao Zexu, Zhou Canquan, Li Jie, Shu Yimin, Liang Xiaoyan, Zhang Mingfang, Zhuang Guanglun
Reproductive Medical Center, First Affiliated Hospital of Sun Yet-Sen University, Guangzhou, China.
Prenat Diagn. 2003 Aug;23(8):646-51. doi: 10.1002/pd.659.
Preimplantation genetic diagnosis (PGD) offers couples at risk for transmitting an inherited disorder the possibility to avoid the need to terminate affected pregnancies. PGD for monogenic diseases is most commonly accomplished by blastomere biopsy from cleavage-stage embryos, followed by PCR-based DNA analysis. However, the molecular heterogeneity of many monogenic diseases requires a diagnostic strategy capable of detecting a range of mutations and compound genotypes. With the above considerations, we developed an accurate and reliable strategy for analysis of beta-globin gene mutations, applicable for PGD for the wide spectrum of beta-thalassemia major mutations in the Chinese population. The strategy involves primer-extension preamplification (PEP), followed by nested PCR and reverse dot blot (RDB) for mutation detection since it facilitates simultaneous analysis of more than one mutation in a single cell. This report describes the application of the strategy in two clinical IVF/PGD cycles at risk for transmitting beta-thalassemia major, which resulted in the first thalassemia-free children born after PGD in China.
植入前基因诊断（PGD）为有遗传疾病传递风险的夫妇提供了避免终止受影响妊娠的可能性。单基因疾病的PGD最常见的方法是从卵裂期胚胎中进行卵裂球活检，随后进行基于PCR的DNA分析。然而，许多单基因疾病的分子异质性需要一种能够检测一系列突变和复合基因型的诊断策略。基于上述考虑，我们开发了一种准确可靠的β-珠蛋白基因突变分析策略，适用于中国人群中广泛的重型β-地中海贫血主要突变的PGD。该策略包括引物延伸预扩增（PEP），随后进行巢式PCR和反向斑点杂交（RDB）以检测突变，因为它有助于在单个细胞中同时分析多个突变。本报告描述了该策略在两个有重型β-地中海贫血传递风险的临床体外受精/PGD周期中的应用，这导致了中国首例PGD后出生的无地中海贫血儿童。
