Well-devised quantification analysis for duplication mutation of Duchenne muscular dystrophy aimed at preimplantation genetic diagnosis

Nakabayashi, Akira; Sueoka, Kou; Tajima, Hiroto; Sakai, Kenji; Sakamoto, Yoshiaki; Katou, Shingo; Yoshimura, Yasunori

doi:10.1007/s10815-007-9111-3

Cited by 3 publications

(3 citation statements)

References 28 publications

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“…In nondeletion DMD cases (duplication, point mutation), linkage analysis and/or gender determination are used for PGD. A quantitative real-time PCR method has been described for PGD of duplication, but no clinical cases have been reported as to our knowledge [7]. …”

Section: Discussionmentioning

confidence: 99%

Preimplantational Genetic Diagnosis and Mutation Detection in a Family with Duplication Mutation of DMD Gene

et al. 2014

Gynecol Obstet Invest

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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.

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Section: Discussionmentioning

confidence: 99%

Preimplantational Genetic Diagnosis and Mutation Detection in a Family with Duplication Mutation of DMD Gene

et al. 2014

Gynecol Obstet Invest

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“…In the next few months, they will be applied to other couples enrolled in the DMD PGD programme. For families showing duplications, a quantitative real-time PCR method was described but no clinical case was reported (Nakabayashi et al, 2007). However, the time and effort necessary to optimize the quantification assays, as advocated by the authors, make haplotype analysis a more attractive alternative.…”

Section: Sequencementioning

confidence: 99%

Rapid and powerful decaplex and dodecaplex PGD protocols for Duchenne muscular dystrophy

Girardet

Fernández

Claustres

2009

Reproductive BioMedicine Online

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Duchenne muscular dystrophy (DMD) is a common childhood lethal X-linked recessive disorder, resulting from deletions, duplications and point mutations in the dystrophin gene. Single-cell protocols for preimplantation genetic diagnosis (PGD) still remain challenging due to the enormous size of the gene and the high risk of intragenic recombination, limitations that often lead to sex determination and selection of female embryos. This study describes direct and rapid decaplex and dodecaplex polymerase chain reaction protocols enabling the analysis of five or seven exons and four microsatellite markers scattered along the dystrophin gene, chosen to be located in the two deletion hotspots, and the analysis of amelogenin sequences for gender determination. The dodecaplex protocol may be applied to most of the couples requesting PGD for DMD in whom the female partner is a carrier of a deletion. This generic approach will allow prompt response to the PGD referrals by reducing the pre-clinical PGD work-up. It was successfully applied in three DMD families, resulting in the birth of a girl as well as in a healthy ongoing pregnancy.

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“…The disease-causing mutation might be passed to offspring and 50% of normal male embryos would be discarded [9]. Later, polymerase chain reaction (PCR) methods, such as multiplex PCR and triplex nested PCR, were applied for direct mutation detection [10][11][12][13]. PCR methods usually divide embryos into two groups, affected and unaffected, while heterozygous carriers would be identified as unaffected.…”

Section: Introductionmentioning

confidence: 99%

Clinical application of an NGS-based method in the preimplantation genetic testing for Duchenne muscular dystrophy

Ren

Lian

Yan

et al. 2021

J Assist Reprod Genet

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Purpose To determine whether next-generation sequencing (NGS) could be used to directly detect different mutations of Duchenne muscular dystrophy (DMD) during preimplantation genetic testing (PGT). Methods From Sep. 2016 to Aug. 2018, a total of six couples participated in this study. Four cases carried DMD exon deletions and two carried exon duplications. Trophectoderm cells were biopsied at day 5 or 6 and NGS was used in the genetic testing of the biopsied cells after whole-genome amplification. We developed a new method-DIRected Embryonic Cell Testing of Exon Deletion/Duplication (DIRECTED) to directly detect the single-gene mutation by NGS. Linage analysis based on singlenucleotide polymorphism (SNP) was used to validate the results from DIRECTED. Results In the four deletion cases, DIRECTED was used to detect DMD exon deletion in 16 biopsied embryos. All DIRECTED results were consistent with linkage analysis, indicating this method was reliable in detecting deletions around 1 Mb. In the two cases carrying exon duplications, no blastocyst was obtained for biopsy. Nonetheless, preliminary experiment results suggested that DIRECTED could also be used for direct detection of exon duplications in embryos. Conclusions Exon deletions or duplications in DMD of preimplantation embryos could be detected directly by NGS-based methods during PGT.

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Well-devised quantification analysis for duplication mutation of Duchenne muscular dystrophy aimed at preimplantation genetic diagnosis

Cited by 3 publications

References 28 publications

Preimplantational Genetic Diagnosis and Mutation Detection in a Family with Duplication Mutation of DMD Gene

Preimplantational Genetic Diagnosis and Mutation Detection in a Family with Duplication Mutation of DMD Gene

Rapid and powerful decaplex and dodecaplex PGD protocols for Duchenne muscular dystrophy

Clinical application of an NGS-based method in the preimplantation genetic testing for Duchenne muscular dystrophy

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