遗传 ›› 1998, Vol. 20 ›› Issue (1): 1-6.

• 论文 • 上一篇    下一篇

73例中国人血友病甲基因突变的分析

刘建湘; 张宇舟; 王鸿利; 黄秋花; 曹文俊; 王学锋; 邵慧珍; 王振义; 陈竺; 黄薇 LIU Jian-Xiang;ZHANG Yu-Zhou;WANG Hong-Li;HUANG Qiu-Hua;CAO Wen-Jun;WANG Xue-Feng;SHAO Hui-Zhen;WANG Zhen-Yi;CHEN Zhu;HANG Wei   

  1. 上海第二医科大学附属瑞金医院, 上海200025 Shanghai Institute of Hematology,Rui Jin Hospital,Shanghai Second Medical University,Shanghai 200025
  • 收稿日期:1900-01-01 出版日期:1998-02-10 发布日期:1998-02-10

Characterizatin of Genetic Defects in 73 Cases of Haemophilia A of Chinese Origin

  • Received:1900-01-01 Online:1998-02-10 Published:1998-02-10

摘要: 我们用Southern blotting、PCR、变性梯度凝胶电泳(DGGE)和DNA测序等方法对73例血友病甲患者(经上海瑞金医院测定血浆FVⅢ:C和vWF:Ag诊断,其中无亲缘关系患者65例,按FVⅢ:C水平分为轻、中、重三型。FVⅢ:C< 2%为重型,共47例;FVⅢ:C 2%-5%为中型,共9例;FVⅢ:C5%-25%为轻型,共1 7例)进行FVⅢ基因突变检测。共检出内含子22倒位23例,均为重型,约占重型的49%,与国外报道相似。余下50例(其中无亲缘关系者45)用PCR-DGG E分析所有外显子及其侧翼内含子序列,发现异常条带则进行DNA测序。在17例患者中检出突变13种,其中无义突变5种,均为重型;错义突变6种,除1例外都是轻中型;小缺失2例,都是重型;其中,AA466Lys(AAG)-Thr(ACG)、719Tyr(TAC)-Stop(TAG)、AA826 Asp(GAC)-Glu(GAA)、312Ile(ATC)-xxC及AA1551-1552del(AGAA)为新发现的突变。有亲缘关系的患者都有相同的基因突变,而在无亲缘关系患者未发现相同突变。基因突变与临床表现基本相符。
Abstract:We use Southern blotting,PCR,denaturing gradient gel electrophoresis(DGGE)and DNA sequencing to detect gene mutations of haemophilia A in Chinese population.73 cases(47 severe)(FVIII:C<2%),9 moderate(FVIII:C 2%~5%),17 mild(FVIII:C 5%~25%)of haemophilia A were first screened with Southern blotting,23 were found to be the intron 22 inversion type,all being severe cases.The remaining 50 cases without intron 22 in version were examined with PCR-DGGE.Genomic DNA were amplified using GC-clamped primers covering all the exons and all flanking intron regions.Abnormal bands were sequenced.13 different mutations were identified,including 5 nonsense mutations,6 missense mutations and 2 small deletions.5 mutations,AA466Lys(AAG)-Thr(ACG),AA719Tyr(TAC)-Stop(TAG),AA826Asp(GAC)-Glu(GAA),AA312Ile(ATC)-xxC and AA1551-1552del(AGAA)have not been reported before.Generally the genetic defects correspond to the clinical conditions.

关键词: DGGE, 变性梯度凝胶电泳, Southern blotting, 基因突变, 血友病甲, EXT基因, 遗传性多发性外生性骨疣病, 连锁分析, 二核苷酸重复

Key words: Hereditary multiple exostoses EXT gene Short-tandem-repeat Linkahe analysis