Tags

Type your tag names separated by a space and hit enter

Simplified PGD of common determinants of haemoglobin Bart's hydrops fetalis syndrome using multiplex-microsatellite PCR.
Reprod Biomed Online. 2010 Nov; 21(5):642-8.RB

Abstract

The high incidence of double-gene deletions in α-thalassaemia increases the risk of having pregnancies with homozygous α(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 α-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 α-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. Alpha-thalassaemia is one of the most common inheritable disorders worldwide. It is a blood disorder that, in its lethal form caused by deletion of all four copies of the α-globin gene, results in the demise of the affected fetus, a condition referred to as haemoglobin (Hb) Bart's hydrops fetalis syndrome. Preimplantation genetic diagnosis (PGD) has played an important role in preventing such cases. Current PGD protocols for deletional α-thalassaemia utilize a strategy called gap-PCR, which requires the different assays for different deletion types. We have developed a universal PGD assay applicable to all common deletional determinants of Hb Bart's hydrops fetalis syndrome based on microsatellite marker analysis. 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. Forty-five embryos were analysed in total. Three pregnancies were achieved from three couples, with the births of two healthy babies and one pregnancy still ongoing. We have successfully adapted our earlier protocol and developed a simple and reliable single cell assay applicable to PGD of Hb Bart's hydrops fetalis syndrome regardless of the type of deletion.

Authors+Show Affiliations

Preimplantation Genetic Diagnosis Center, University Children's Medical Institute, Department of Pediatrics, Yong Loo Lin School of Medicine, National University Health System, Singapore 119074, Singapore.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20864413

Citation

Wang, Wen, et al. "Simplified PGD of Common Determinants of Haemoglobin Bart's Hydrops Fetalis Syndrome Using Multiplex-microsatellite PCR." Reproductive Biomedicine Online, vol. 21, no. 5, 2010, pp. 642-8.
Wang W, Yap CH, Loh SF, et al. Simplified PGD of common determinants of haemoglobin Bart's hydrops fetalis syndrome using multiplex-microsatellite PCR. Reprod Biomed Online. 2010;21(5):642-8.
Wang, W., Yap, C. H., Loh, S. F., Tan, A. S., Lim, M. N., Prasath, E. B., Chan, M. L., Tan, W. C., Jiang, B., Yeo, G. H., Mathew, J., Ho, A., Ho, S. S., Wong, P. C., Choolani, M. A., & Chong, S. S. (2010). Simplified PGD of common determinants of haemoglobin Bart's hydrops fetalis syndrome using multiplex-microsatellite PCR. Reproductive Biomedicine Online, 21(5), 642-8. https://doi.org/10.1016/j.rbmo.2010.06.021
Wang W, et al. Simplified PGD of Common Determinants of Haemoglobin Bart's Hydrops Fetalis Syndrome Using Multiplex-microsatellite PCR. Reprod Biomed Online. 2010;21(5):642-8. PubMed PMID: 20864413.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Simplified PGD of common determinants of haemoglobin Bart's hydrops fetalis syndrome using multiplex-microsatellite PCR. AU - Wang,Wen, AU - Yap,Christine H A, AU - Loh,Seong Feei, AU - Tan,Arnold S C, AU - Lim,Mui Nee, AU - Prasath,Ethiraj B, AU - Chan,Melinda L H, AU - Tan,Wei Chin, AU - Jiang,Boran, AU - Yeo,Gare Hoon, AU - Mathew,Joyce, AU - Ho,Angela, AU - Ho,Sherry S Y, AU - Wong,Peng Cheang, AU - Choolani,Mahesh A, AU - Chong,Samuel S, Y1 - 2010/06/19/ PY - 2010/01/11/received PY - 2010/06/03/revised PY - 2010/06/08/accepted PY - 2010/9/25/entrez PY - 2010/9/25/pubmed PY - 2011/2/18/medline SP - 642 EP - 8 JF - Reproductive biomedicine online JO - Reprod. Biomed. Online VL - 21 IS - 5 N2 - The high incidence of double-gene deletions in α-thalassaemia increases the risk of having pregnancies with homozygous α(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 α-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 α-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. Alpha-thalassaemia is one of the most common inheritable disorders worldwide. It is a blood disorder that, in its lethal form caused by deletion of all four copies of the α-globin gene, results in the demise of the affected fetus, a condition referred to as haemoglobin (Hb) Bart's hydrops fetalis syndrome. Preimplantation genetic diagnosis (PGD) has played an important role in preventing such cases. Current PGD protocols for deletional α-thalassaemia utilize a strategy called gap-PCR, which requires the different assays for different deletion types. We have developed a universal PGD assay applicable to all common deletional determinants of Hb Bart's hydrops fetalis syndrome based on microsatellite marker analysis. 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. Forty-five embryos were analysed in total. Three pregnancies were achieved from three couples, with the births of two healthy babies and one pregnancy still ongoing. We have successfully adapted our earlier protocol and developed a simple and reliable single cell assay applicable to PGD of Hb Bart's hydrops fetalis syndrome regardless of the type of deletion. SN - 1472-6491 UR - https://www.unboundmedicine.com/medline/citation/20864413/Simplified_PGD_of_common_determinants_of_haemoglobin_Bart's_hydrops_fetalis_syndrome_using_multiplex_microsatellite_PCR_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1472-6483(10)00402-5 DB - PRIME DP - Unbound Medicine ER -