Rapid Screening for Japanese Dysferlinopathy by Fluorescent Primer Extension
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- Hayashi Saori
- Department of Neurology, Kawasaki Medical School
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- Ohsawa Yutaka
- Department of Neurology, Kawasaki Medical School
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- Takahashi Toshiaki
- Department of Neurology, Nishitaga National Hospital
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- Suzuki Naoki
- Department of Neurology, Tohoku University School of Medicine
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- Okada Tadashi
- Department of Neurology, Kawasaki Medical School
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- Rikimaru Mitsue
- Department of Neurology, Kawasaki Medical School
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- Murakami Tatsufumi
- Department of Neurology, Kawasaki Medical School
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- Aoki Masashi
- Department of Neurology, Tohoku University School of Medicine
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- Sunada Yoshihide
- Department of Neurology, Kawasaki Medical School
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Objective Mutations in the dysferlin gene cause limb-girdle muscular dystrophy (LGMD) 2B and Miyoshi myopathy (MM), which are collectively named dysferlinopathy. Dysferlinopathy is the most frequent type of LGMD in the Japanese population. Molecular genetic analysis is essential for the diagnosis of dysferlinopathy because of its variable immunohistochemical patterns of biopsied muscles, including patterns similar to normal controls. The analysis of the entire dysferlin gene however, is time-consuming and laborious; therefore a simple and rapid screening method to detect hot spot mutations in the dysferlin gene is essential for the diagnosis of dysferlinopathy.<br> Methods We previously showed that 4 mutations, c.937+1G>A, c.1566C>G, c.2997G>T and c.3373delG account for 50% of all the mutations identified in Japanese dysferlinopathy patients. We performed a one-tube multiplex PCR, followed by extension of primers for each mutation with a fluorescence-labeled dideoxynucleotide to screen the 4 hot spot mutations.<br> Results The multiplex primer-extension reaction was developed on samples of known mutations. The extension products were represented as 4 different peaks that corresponded to a mutated nucleotide on electropherogram. Using the developed screening method, we were able to detect mutations in these hot spots in 3 samples out of 8 clinically suspected LGMD2B/MM patients in only approximately 8 hours. These 3 cases were definitely diagnosed as LGMD2B/MM by exonic sequencing.<br> Conclusion We have developed a simple and rapid screening method which could facilitate the definitive diagnosis of dysferlinopathy, contributing to an understanding of the genotype-phenotype correlations for dysferlinopathy.<br>
収録刊行物
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- Internal Medicine
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Internal Medicine 49 (24), 2693-2696, 2010
一般社団法人 日本内科学会
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詳細情報 詳細情報について
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- CRID
- 1390282679847362304
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- NII論文ID
- 130000413429
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- ISSN
- 13497235
- 09182918
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- 本文言語コード
- en
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- データソース種別
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- JaLC
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