应用新型多重荧光 PCR 技术对α-地中海贫血-SEA 进行植入前遗传学诊断。
Preimplantation genetic diagnosis of alpha-thalassemia-SEA using novel multiplex fluorescent PCR.
机构信息
Preimplantation Genetic Diagnosis Center, Department of Obstetrics and Gynaecology, Faculty of Medicine, Chiang Mai University, 110 Intawaroros Road, Sripoom, Mueang, Chiang Mai, 50200, Thailand.
出版信息
J Assist Reprod Genet. 2012 Jan;29(1):95-102. doi: 10.1007/s10815-011-9641-6. Epub 2011 Oct 1.
PURPOSE
Preimplantation genetic diagnosis (PGD) is an alternative to prenatal diagnosis (PND) giving couples at risk a chance to start a pregnancy with a disease-free baby. This study aimed to develop a new PGD protocol for alpha-thalassemia(-SEA) mutation, the commonest Mendelian disorder.
PATIENTS AND METHODS
Multiplex fluorescent PCR was employed for mutation, contamination and linkage analysis. A couple experienced termination of pregnancy following positive PND decided to join the project.
RESULTS
Novel primers for alpha-thalassemia(-SEA) mutation amplifying 5 DNA fragments were developed. Two PGD cycles were performed, resulting in an un-affected baby. PND confirmed the heterozygous result. From 24 embryos, 87.5% of affected genotype were of best quality compared to 0% and 18.2% of those with normal and heterozygous, respectively.
CONCLUSIONS
A novel PCR protocol for the common alpha-thalassemia(-SEA) mutation is reported. This test should be widely applicable. Interestingly, a potential effect of alpha-thalassemia(-SEA) mutation on preimplantation embryonic development was noticed.
目的
植入前遗传学诊断（PGD）是一种替代产前诊断（PND）的方法，为有风险的夫妇提供了一个机会，让他们能够怀上一个没有疾病的婴儿。本研究旨在为最常见的孟德尔疾病--缺失型α-地中海贫血（-SEA）突变，开发一种新的 PGD 方案。
患者与方法
采用多重荧光 PCR 进行突变、污染和连锁分析。一对夫妇在 PND 阳性后决定终止妊娠，并决定参与该项目。
结果
开发了用于缺失型α-地中海贫血（-SEA）突变扩增 5 个 DNA 片段的新型引物。进行了两个 PGD 周期，最终生下了一个未受影响的婴儿。PND 证实了杂合子的结果。在 24 个胚胎中，87.5%的受影响基因型的胚胎质量最好，而正常和杂合子的胚胎质量分别为 0%和 18.2%。
结论
报道了一种用于常见缺失型α-地中海贫血（-SEA）突变的新型 PCR 方案。该检测方法应该具有广泛的适用性。有趣的是，我们注意到缺失型α-地中海贫血（-SEA）突变对胚胎植入前发育可能有潜在影响。
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