【作者】 石西南；

【导师】 褚嘉祐； 杨昭庆；

【作者基本信息】 昆明医学院 ， 生物化学与分子生物学， 2011， 硕士

【摘要】 β-地中海贫血是p珠蛋白基因突变或部分缺失导致p珠蛋白肽链合成速率降低或缺如,导致红细胞渗透脆性减少和血中HbA2水平升高为特征的遗传性溶血性贫血病。β-地中海贫血主要有两类：完全不能合成β链的称为βp0地贫,能部分合成β链(约为正常的5%-30%)的称为β+地贫。地中海、非洲、东南亚、印度次大陆以及我国西南、华南地区是地中海贫血高发区域。β-地中海贫血在云南少数民族傣族和景颇族中高发,是常见的遗传性血液病之一,有文献报道云南德宏傣族、景颇族发病率为10.0%。其分子病理具有高度异质性,主要为β珠蛋白基因点突变、缺失或插入。目前全世界已报道了约200多种HBB基因突变,在中国人群中已报道有34种,我国β-地中海贫血突变常见类型为CD41-42、IVS-Ⅱ-654、CD17、TATABbox-28.CD71-72和TATAbox-29等。研究表明：β-地中海贫血的基因突变分布具有明显的种族特征和地域差异,云南西双版纳傣族居住在中国西南亚热带湿热地区,其地理位置独特,国内外对本地区β-地中海贫血的报道还不多见。本研究采用了DNA序列分析的方法分析了云南傣族儿童中β-地中海贫血β珠蛋白基因变异分布特点,并建立检测相应常见变异位点的快速筛查方法。结果显示：209例样本中共发现9个变异位点,按频率由高到低分别是CD2 T>C(50.72%)、CD26 G>A(35.41%)、CD17 A>T(12.92%)、IVS-Ⅱ-17 C>A(12.44%)、IVS-Ⅱ-16 G>C(11.96%).IVS-Ⅰ-30A>G(9.57%).CD6T>A (9.09%)、CD41-42 (-TCTT)(7.18%)和CD5 T>A(1.92%),有157人检测出基因变异位点,占总人数的73.21%。引入错配碱基的ARMS引物,在一定的的退火温度条件下能对CD2、CD26和CD17三个常见变异位点进行特异筛查。本实验结果显示云南西双版纳傣族β-地中海贫血人群中HBB基因的变异位点有其自身的特点,与其他地区有所不同。ARMS检测云南傣族β-地中海贫血基因变异是一种准确有效并简便经济的方法。更多还原

【Abstract】 Theβ-thalassaemia is hereditary autosomal disorders with decreased(β+) or absent(β0) (5%-30%)β-globin chain synthesis.The most common genetic defects inβ-thalassaemia are caused by point mutations or small deletions or insertions in HBB gene, which caused a substantial reducion of P-chain synthesis and a distinct haematological phenotye with hypochromic microcytic red blood cells,decreased osmotic fragility and characteristically raised levels of HbA2 in heterozygots. Theβ-thalassaemia is a highly heterogeneous inherited disorder distributed worldwide, which is found in Mediterranean,Africa, southeast Asia, Indian subcontinent and Southwest(Yunnan Dai,Jingpo)(20.00%), South of china. More than 200 such mutations are known at present.34 kinds of mutations are reported in China at present. CD41-42,IVS-Ⅱ-654,CD17,TATABbox-28,CD71-72和TATAbox-29 are common mutations. The distribution characteristics of HBB gene variation in each ethnic population are different, which have their own cluster of common mutations. Xishuangbanna-Dai of Yunnan inhabits Southwest of China, who locates Subtropicality. Only a few mutation of HBB gene were report in this place. In this study, the distribution characterization of variation in HBB gene ofβ-thalassemia in Dai minority group in Yunnan province was analyzed by DNA sequencing. In addition, to estnablish a rapid and economic method for screening mutation inβ-thalassemia by optimizing ARMS technique.Date showed:among the 209 samples, there are 9 variations detected, CD2 T>C (50.72%),CD26 G>A (35.41%),CD17 A>T (12.92%) JVS-Ⅱ-17 C>A (12.44%), IVS-Ⅱ-16 G>C (11.96%), IVS-Ⅰ-30 A>G (9.57%), CD6 T>A (9.09%), CD41-42 (-TCTT) (7.18%) and CD5 T>A (1.92%). There were 153 of 209 participants (73.21%) carrying HBB gene variation. Three common mutations, CD2, CD26 and CD17 can be efficiently detected by combining annealing condition and ARMS mismatch primers. The distribution of HBB gene variation in the Dai minority group inYunnan province were different from that of other groups in china. ARMS is a effective, convenient and economical technique for rapid detection of gene variation inβ-thalassemia.更多还原