Simplified PGD of common determinants of haemoglobin Bart's hydrops fetalis syndrome using multiplex-microsatellite PCR

REPRODUCTIVE BIOMEDICINE ONLINE

卷:21

Article

Wang, Wen¹  Yap, Christine H. A.²  Loh, Seong Feei³  Tan, Arnold S. C.¹  Lim, Mui Nee²  Prasath, Ethiraj B.³  Chan, Melinda L. H.³  Tan, Wei Chin¹  Jiang, Boran¹  Yeo, Gare Hoon¹  Ho, Angela⁴  Chong, Samuel S.¹ 

[1] Natl Univ Hlth Syst, Univ Childrens Med Inst, Preimplantat Genet Diag Ctr, Dept Pediat,Yong Loo Lin Sch Med, Singapore 119074, Singapore

[2] Singapore Gen Hosp, Dept Obstet & Gynecol, Ctr Assisted Reprod, Singapore 169608, Singapore

[3] KK Womens & Childrens Hosp, KK IVF, Dept Reprod Med, Singapore 229899, Singapore

[4] Ctr Assisted Reprod Private Ltd, Paragon Med Ctr, Singapore 238859, Singapore

关键词: alpha-thalassaemia;    Hb Bart's hydrops fetalis syndrome;    microsatellite marker;    polymerase chain reaction;    preimplantation genetic diagnosis;   

DOI  :  10.1016/j.rbmo.2010.06.021

来源: Elsevier

【 摘 要 】

The high incidence of double-gene deletions in alpha-thalassaemia increases the risk of having pregnancies with homozygous alpha(0)-thalassaemia, the cause of the lethal haemoglobin (Hb) Bart's hydrops fetalis syndrome. Preimplantation genetic diagnosis (PGD) has played an important role in preventing such cases. However, the current gap-PCR based PGD protocol for deletional alpha-thalassaemia requires specific primer design for each specific deletion. A universal PGD assay applicable to all common deletional determinants of Hb Bart's hydrops fetalis syndrome has been developed. Microsatellite markers 16PTEL05 and 16PTEL06 within the alpha-globin gene cluster were co-amplified with a third microsatellite marker outside the affected region in a multiplex-PCR reaction and analysed by capillary electrophoresis. Eight informed couples at risk of having Hb Bart's hydrops fetalis were recruited in this study and all patients underwent standard procedures associated with IVF. A total of 47 embryos were analysed. Three pregnancies were achieved from three couples, with the births of two healthy babies and one ongoing pregnancy. This work has successfully adapted an earlier protocol and developed a simple and reliable single-cell assay applicable to PGD of Hb Bart's hydrops fetalis syndrome regardless of type of deletion. (C) 2010, Reproductive Healthcare Ltd. Published by Elsevier Ltd. AU rights reserved.

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