Next-generation sequencing discloses a nonsense mutation in the dystrophin gene from long preserved dried umbilical cord and low-level somatic mosaicism in the proband mother

Mariko Ikeda, Yasuhiro Takeshima, Tomoko Lee, Masahiro Nishiyama, Hiroyuki Awano, Mariko Yagi, Ai Unzaki, Kandai Nozu, Hisahide Nishio, Masafumi Matsuo, Hiroki Kurahashi, Tatsushi Toda, Ichiro Morioka, Kazumoto Iijima

研究成果: Article

5 引用 (Scopus)

抄録

Duchene muscular dystrophy (DMD) is a progressive muscle wasting disease, caused by mutations in the dystrophin (DMD) on the X chromosome. One-third of patients are estimated to have de novo mutations. To provide in-depth genetic counseling, the comprehensive identification of mutations is mandatory. However, many DMD patients did not undergo genetic diagnosis because detailed genetic diagnosis was not available or their mutational types were difficult to identify. Here we report the genetic testing of a sporadic DMD boy, who died >20 years previously. Dried umbilical cord preserved for 38 years was the only available source of genomic DNA. Although the genomic DNA was severely degraded, multiplex ligation-dependent probe amplification analysis was performed but no gross mutations found. Sanger sequencing was attempted but not conclusive. Next-generation sequencing (NGS) was performed by controlling the tagmentation during library preparation. A nonsense mutation in DMD (p.Arg2095∗) was clearly identified in the proband. Consequently, the identical mutation was detected as an 11% mosaic mutation from his healthy mother. Finally, the proband's sister was diagnosed as a non-carrier of the mutation. Thus using NGS we have identified a pathogenic DMD mutation from degraded DNA and low-level somatic mosaicism, which would have been overlooked using Sanger sequencing.

元の言語English
ページ(範囲)351-355
ページ数5
ジャーナルJournal of Human Genetics
61
発行部数4
DOI
出版物ステータスPublished - 01-04-2016

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Dystrophin
Mosaicism
Nonsense Codon
Umbilical Cord
Muscular Dystrophies
Mothers
Mutation
Genes
DNA
Wasting Syndrome
Multiplex Polymerase Chain Reaction
Genetic Counseling
Genetic Testing
X Chromosome
Libraries
Siblings
Muscles

All Science Journal Classification (ASJC) codes

  • Genetics
  • Genetics(clinical)

これを引用

Ikeda, Mariko ; Takeshima, Yasuhiro ; Lee, Tomoko ; Nishiyama, Masahiro ; Awano, Hiroyuki ; Yagi, Mariko ; Unzaki, Ai ; Nozu, Kandai ; Nishio, Hisahide ; Matsuo, Masafumi ; Kurahashi, Hiroki ; Toda, Tatsushi ; Morioka, Ichiro ; Iijima, Kazumoto. / Next-generation sequencing discloses a nonsense mutation in the dystrophin gene from long preserved dried umbilical cord and low-level somatic mosaicism in the proband mother. :: Journal of Human Genetics. 2016 ; 巻 61, 番号 4. pp. 351-355.
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abstract = "Duchene muscular dystrophy (DMD) is a progressive muscle wasting disease, caused by mutations in the dystrophin (DMD) on the X chromosome. One-third of patients are estimated to have de novo mutations. To provide in-depth genetic counseling, the comprehensive identification of mutations is mandatory. However, many DMD patients did not undergo genetic diagnosis because detailed genetic diagnosis was not available or their mutational types were difficult to identify. Here we report the genetic testing of a sporadic DMD boy, who died >20 years previously. Dried umbilical cord preserved for 38 years was the only available source of genomic DNA. Although the genomic DNA was severely degraded, multiplex ligation-dependent probe amplification analysis was performed but no gross mutations found. Sanger sequencing was attempted but not conclusive. Next-generation sequencing (NGS) was performed by controlling the tagmentation during library preparation. A nonsense mutation in DMD (p.Arg2095∗) was clearly identified in the proband. Consequently, the identical mutation was detected as an 11{\%} mosaic mutation from his healthy mother. Finally, the proband's sister was diagnosed as a non-carrier of the mutation. Thus using NGS we have identified a pathogenic DMD mutation from degraded DNA and low-level somatic mosaicism, which would have been overlooked using Sanger sequencing.",
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Ikeda, M, Takeshima, Y, Lee, T, Nishiyama, M, Awano, H, Yagi, M, Unzaki, A, Nozu, K, Nishio, H, Matsuo, M, Kurahashi, H, Toda, T, Morioka, I & Iijima, K 2016, 'Next-generation sequencing discloses a nonsense mutation in the dystrophin gene from long preserved dried umbilical cord and low-level somatic mosaicism in the proband mother', Journal of Human Genetics, 巻. 61, 番号 4, pp. 351-355. https://doi.org/10.1038/jhg.2015.157

Next-generation sequencing discloses a nonsense mutation in the dystrophin gene from long preserved dried umbilical cord and low-level somatic mosaicism in the proband mother. / Ikeda, Mariko; Takeshima, Yasuhiro; Lee, Tomoko; Nishiyama, Masahiro; Awano, Hiroyuki; Yagi, Mariko; Unzaki, Ai; Nozu, Kandai; Nishio, Hisahide; Matsuo, Masafumi; Kurahashi, Hiroki; Toda, Tatsushi; Morioka, Ichiro; Iijima, Kazumoto.

:: Journal of Human Genetics, 巻 61, 番号 4, 01.04.2016, p. 351-355.

研究成果: Article

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AU - Ikeda, Mariko

AU - Takeshima, Yasuhiro

AU - Lee, Tomoko

AU - Nishiyama, Masahiro

AU - Awano, Hiroyuki

AU - Yagi, Mariko

AU - Unzaki, Ai

AU - Nozu, Kandai

AU - Nishio, Hisahide

AU - Matsuo, Masafumi

AU - Kurahashi, Hiroki

AU - Toda, Tatsushi

AU - Morioka, Ichiro

AU - Iijima, Kazumoto

PY - 2016/4/1

Y1 - 2016/4/1

N2 - Duchene muscular dystrophy (DMD) is a progressive muscle wasting disease, caused by mutations in the dystrophin (DMD) on the X chromosome. One-third of patients are estimated to have de novo mutations. To provide in-depth genetic counseling, the comprehensive identification of mutations is mandatory. However, many DMD patients did not undergo genetic diagnosis because detailed genetic diagnosis was not available or their mutational types were difficult to identify. Here we report the genetic testing of a sporadic DMD boy, who died >20 years previously. Dried umbilical cord preserved for 38 years was the only available source of genomic DNA. Although the genomic DNA was severely degraded, multiplex ligation-dependent probe amplification analysis was performed but no gross mutations found. Sanger sequencing was attempted but not conclusive. Next-generation sequencing (NGS) was performed by controlling the tagmentation during library preparation. A nonsense mutation in DMD (p.Arg2095∗) was clearly identified in the proband. Consequently, the identical mutation was detected as an 11% mosaic mutation from his healthy mother. Finally, the proband's sister was diagnosed as a non-carrier of the mutation. Thus using NGS we have identified a pathogenic DMD mutation from degraded DNA and low-level somatic mosaicism, which would have been overlooked using Sanger sequencing.

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