Single-tube nonaplex microsatellite PCR panel for preimplantation genetic diagnosis of Hb Bart's hydrops fetalis syndrome.

Autor: Chen M; Yong Loo Lin School of Medicine, Department of Pediatrics, National University of Singapore, Singapore., Chan JK; KKIVF Center, Department of Reproductive Medicine, KK Women's and Children's Hospital, Singapore., Nadarajah S; KKIVF Center, Department of Reproductive Medicine, KK Women's and Children's Hospital, Singapore., Tan AS; Yong Loo Lin School of Medicine, Department of Pediatrics, National University of Singapore, Singapore.; Preimplantation Genetic Diagnosis Center, Khoo Teck Puat - National University Children's Medical Institute, National University Health System, Singapore., Chan ML; KKIVF Center, Department of Reproductive Medicine, KK Women's and Children's Hospital, Singapore., Mathew J; Clinic for Human Reproduction, Department of Obstetrics and Gynecology, National University Hospital, Singapore., Saw EE; Preimplantation Genetic Diagnosis Center, Khoo Teck Puat - National University Children's Medical Institute, National University Health System, Singapore., Lim C; KKIVF Center, Department of Reproductive Medicine, KK Women's and Children's Hospital, Singapore., Wong W; KKIVF Center, Department of Reproductive Medicine, KK Women's and Children's Hospital, Singapore., Cheah FS; Preimplantation Genetic Diagnosis Center, Khoo Teck Puat - National University Children's Medical Institute, National University Health System, Singapore., Law HY; Department of Paediatric Medicine, KK Women's and Children's Hospital, Singapore.; Paediatrics Academic Clinical Program, Duke-NUS Graduate Medical School, Singapore., Wong PC; Clinic for Human Reproduction, Department of Obstetrics and Gynecology, National University Hospital, Singapore., Chong SS; Yong Loo Lin School of Medicine, Department of Pediatrics, National University of Singapore, Singapore.; Preimplantation Genetic Diagnosis Center, Khoo Teck Puat - National University Children's Medical Institute, National University Health System, Singapore.; Department of Laboratory Medicine, National University Hospital, Singapore.
Jazyk: angličtina
Zdroj: Prenatal diagnosis [Prenat Diagn] 2015 Jun; Vol. 35 (6), pp. 534-43. Date of Electronic Publication: 2015 Mar 01.
DOI: 10.1002/pd.4568
Abstrakt: Objective: To develop a single-tube multi-marker assay for improved preimplantation genetic diagnosis (PGD) of deletional and/or non-deletional Hb Bart's hydrops fetalis syndrome, providing haplotype confirmation of deletional status, and maximization of linkage informativity.
Methods: We performed in silico mining to identify novel microsatellites within 1 Mb flanking the alpha-globin gene cluster, and optimized a single-tube assay combining detection of α(0) -thalassemia deletions with multi-marker linkage analysis. We performed validation on 100 single cells prior to clinical PGD application.
Results: Of 42 markers encompassing the α-globin gene cluster that were identified in silico, 9 were highly polymorphic (0.68 ≤ polymorphism information content ≤ 0.92; 0.66 ≤ Ho  ≤ 0.90; 10 ≤ alleles ≤ 35) and optimized to co-amplify directly from a single cell. A validation analysis of 100 single lymphoblasts yielded 100% amplification success for all markers, and individual marker allele drop-out (ADO) rates of 0-5%. Clinical application of the assay in PGD for Hb Bart's (2 cases/cycles) resulted in a twin pregnancy and healthy live birth of two baby girls.
Conclusions: This single-tube nonaplex microsatellite PCR panel can be applied directly to PGD of most deletional Hb Bart's without the need for deletion-specific customization, and to linkage-based PGD of non-deletional Hb Bart's.
(© 2015 John Wiley & Sons, Ltd.)
Databáze: MEDLINE
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