title: Preimplantational genetic diagnosis and mutation detection in a family with duplication mutation of DMD gene.
abstract: Duchenne muscular dystrophy (DMD) is an X-linked recessive neuromuscular disease caused by mutation in the DMD gene. A 38-year-old woman was referred to our hospital with her son who was diagnosed with DMD. Multiplex PCR failed to detect DMD mutations in the affected child. The female carrier underwent preimplantation genetic diagnosis by linkage analysis and gender determination. Eight embryos were biopsied after in vitro fertilization. Two healthy embryos determined both as females (E1 and E3) were transferred. Although the paternal allele was absent in E3, it was considered to be a result of allele dropout for the STR-49 marker. Surprisingly, a female and a male baby were delivered at 38 gestational weeks, suggesting that E3 was a male embryo with the allele dropout occurring at the SRY gene. Exon 2 duplication was detected in the DMD patient and the carrier mother using next-generation sequencing and multiple ligation-dependent probe amplification. Next, we verified the duplication of exon 2 by real-time PCR, using a special primer at 3' of intron 1, very close to exon 2. Finally, we confirmed that both newborns inherited the normal allele, using quantitative real-time PCR and linkage analysis.
authors.affiliation: Department of Reproductive Endocrinology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, China.
authors.forename: Yinghui
authors.initials: Y
authors.lastname: Ye
authors.forename: Ping
authors.initials: P
authors.lastname: Yu
authors.forename: Jing
authors.initials: J
authors.lastname: Yong
authors.forename: Ting
authors.initials: T
authors.lastname: Zhang
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Wei
authors.forename: Ming
authors.initials: M
authors.lastname: Qi
authors.forename: Fan
authors.initials: F
authors.lastname: Jin
mesh_terms: D000328:Adult; D000483:Alleles; D002648:Child; D004252:DNA Mutational Analysis / Q000379:methods; D004624:Embryo Transfer; D005091:Exons; D005260:Female; D005307:Fertilization in Vitro; D020440:Gene Duplication; D006580:Genetic Carrier Screening; D008040:Genetic Linkage; D006801:Humans; D008297:Male; D060885:Multiplex Polymerase Chain Reaction; D020388:Muscular Dystrophy, Duchenne / Q000175:diagnosis / Q000235:genetics*; D019836:Preimplantation Diagnosis / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction