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フルテキストURLA050066000105.pdf ( 3.0MB ) 公開日 2019-09-20
タイトルDiagnosis of α-thalassemia using droplet digital PCR
作成者Amao, Yuki
Yamashiro, Yasuhiro
Hattori, Yukio
Ogata, Shizuka
Ohto, Fukumi
Mella, Ferania
Kimoto, Masafumi
Mori, Kentaro
Suehiro, Yutaka
Yamasaki, Takahiro
作成者ヨミアマオ, ユウキ
ヤマシロ, ヤスヒロ
ハットリ, ユキオ
オガタ, シズカ
オオトウ, フクミ
キモト, マサフミ
モリ, ケンタロウ
スエヒロ, ユタカ
ヤマサキ, タカヒロ
作成者別表記天尾, 優希
山城, 安啓
服部, 幸夫
緒方, 静
大峠, ふくみ
木本, 真史
森, 健太郎
末廣, 寛
山﨑, 隆弘
作成者所属山口大学大学院医学系研究科
内容記述(抄録等)Most α-thalassemia occurs due to a large deletion in the α-globin gene. Common α-thalassemia with known mutations, such as Southeast Asian (SEA) type is readily diagnosed by Gap-PCR. However, there are many unknown mutations in α-thalassemia that need dosage or copy number assessment of the α-globin gene for diagnosis. Unlike β-thalassemia, a quantitative or real-time PCR approach for gene dosage study is not available for α-thalassemia. In real-time PCR, the gene needs to be amplified by two-fold at each cycle for dosage studies, which does not seem to be the case for the α-globin gene. The droplet digital PCR is not affected by amplification efficiency, and accurate quantification or copy number determination (copy/μg DNA) can be obtained. Here, we evaluated droplet digital PCR for detecting α-globin gene deletions. We analyzed DNA from 292 blood samples, including 62 normal samples, 35 heterozygous -α^{3.7}, 19 homozygous -α^{3.7}, 83 SEA type, 17 Filipino (FIL) type, 23 hemoglobin H disease (-α^{3.7}/SEA or FIL), 4α^{CS } /SEA and 49 non-SEA/FIL type α-thalassemia. Results of all α-thalassemia cases conformed to predicted values of copy numbers, except for six non-SEA/FIL α-thalassemias that had no deletion of the α-globin gene, but had deletions in a region containing multispecies conserved sequence R2 (DNase I hypersensitive sites 40) that is located 40 kb upstream of the ζ-globin gene and associates with the regulation of the α-globin gene. The α^{CS } allele or α^{T} has no deletion, and is diagnosed by sequencing. Thus, we show that digital droplet PCR gives accurate copy number of the α-globin gene, and is a reliable tool for determination of α-thalassemia.
本文言語eng
著者キーワードDroplet Digital PCR (ddPCR)
α-thalassemia
quntitative PCR
copy number variation
資料タイプtext
ファイル形式application/pdf
出版者Yamaguchi University School of Medicine
NII資料タイプ紀要論文
ISSN0513-1812
NCIDAA00594272
学内刊行物(紀要等)The bulletin of the Yamaguchi Medical School
掲載誌名The bulletin of the Yamaguchi Medical School
66
1-2
開始ページ37
終了ページ44
発行日2019
著者版/出版社版出版社版
リポジトリIDA050066000105
地域区分山口大学
URIhttp://www.lib.yamaguchi-u.ac.jp/yunoca/handle/A050066000105