The clinical application of NGS-based SNP haplotyping for PGD of Hb H disease. (4th May 2017)

• Record Type: Journal Article
• Authors: Chen, Linjun; Diao, Zhenyu; Xu, Zhipeng; Zhou, Jianjun; Yan, Guijun; Sun, Haixiang
• Is Part Of: Systems biology in reproductive medicine. Volume 63:Number 3(2017:Jun.)
• Page Start: 212
• Page End: 217
• Publication Date: 2017-05-04
• Subjects: Haplotyping -- hemoglobin H -- next-generation sequencing -- preimplantation genetic diagnosis -- single nucleotide polymorphism; Systems biology -- Periodicals; Andrology -- Periodicals; Generative organs, Male -- Diseases -- Periodicals; Biological systems -- Periodicals; Reproductive health -- Periodicals; Human reproduction -- Periodicals; 612.61
• DOI: 10.1080/19396368.2017.1296501
• Languages: English
• ISSNs: 1939-6368

Abstract

ABSTRACT: This study investigated the usefulness of next-generation sequencing (NGS)-based single nucleotide polymorphism (SNP) haplotyping for preimplantation genetic diagnosis (PGD) of hemoglobin H (Hb H) disease. Multiple displacement amplification (MDA) was used for whole genome amplification (WGA) of biopsied trophectoderm (TE) cells. Gap-PCR and NGS-based SNP haplotyping was used to distinguish the two genotypes of -α 3.7 /αα and – SEA /αα for PGD of Hb H disease. One out of the ten blastocysts (B11) was successfully diagnosed as genotype -α 3.7 /αα by Gap-PCR, whereas the others revealed allele dropout (ADO) (B1, B2, B4, B5, B7, B8, B12, and B15) or amplification failure (B10). However, NGS-based SNP haplotyping successfully diagnosed the -α 3.7 /αα and – SEA /αα genotypes from the MDA products of the biopsied TE cells. The haplotyping result showed that B4, B7, B8, B10, B11, B12, and B15 were carriers of the -α 3.7 deletion (-α 3.7 /αα), whereas B1, B2, and B5 were carriers of the – SEA deletion (– SEA /αα). A blastocyst (B11) was transferred into the uterus in a subsequent frozen embryo transfer (FET) cycle after PGD. A healthy infant with a -α 3.7 /αα genotype weighing 2, 800 g was born by cesarean section at the 38 th week of gestation. This result indicates that NGS-based SNP haplotyping is a valid screening tool for the PGD of Hb H disease.