摘要
本文探讨β—地中海贫血人群普查和基因分析的方法。采一次未梢血,完成β—地中海贫血对象调查和基因类型分析:检查项目:红细胞脆性试验(0.32％Nacl溶液,一管法),Hb微量CAM电泳、目测Hb电泳图判断HbA_2是否增高,DEAE微量柱层析法测定HbA_2含量;白细胞部分供提取DNA进行PCR扩增及ASO探针杂交基因分析。红细胞脆性试验阳性及HbA_2含量超过3.5％(疟疾高发区>4％)者为β—地中海贫血对象。将这些对象的末梢血提取DNA,做PCR扩增及ASO探针杂交法基因分析。普查军田乡黎族小学生730人,筛出β—地贫对象49人。随机用PCR—ASO法检测28例β—地贫对象,检出β—地CD41/d2(—TCTT)基因型杂合子27例,未查明1例,符合率达96％以上。非β—地中海贫血6例经PCR—ASO法检测均非β—地贫基因变异。本文对β—地中海贫血普查结合PCR—ASO基因诊断的方法进行了详细讨论。
An easy and simple method was described here for both the survey of β—Thal and further analyzing the spectrum of β—Thal mutations by collecting human peripheral blood for once only. The qualitative osmostic fragility test (QOFT screening in 0. 32％ NaCl) and Hb micro—CAM eletrophoresis were used for screening out the cases of β—Thal first, after the DNA was isolated from the pellet of white cells, then analyzed by the raised HbA_2 can be visualzed on the Hb electropherogram and if the amount of it is over 3.5％ (or 4. 0％ in malaria current zone) it can be detected by the method of DEAE microcolumn chromatography. Totaled 730 primary students of the Li nationality in Juntian village, Hainan Island, were investigated, and 49 cases of β—Thal were screened out by the QOFT and the Hb electropherogram. Among the 49 cases 28 were randomly chosen for further analyzed by the PCR—ASO method. 27 cases of β—Thal genotype (CD41/42,—TCTT) were found, one of them was unidentified, and the credible rate of screening out β—Thal by the QOGT and the Hb electropherogram was over 96％. The another 6 cases of non—β—Thal were tested by the QOFT and Hb electropherogram it were further proceed not of β—Thal genotype by the PCR—ASO method. A detail discusion was gaven in this paper about the survey of the β—Thal with the PCR—ASO gene diagnosed bythe PCR—ASO method.
出处
《海南大学学报（自然科学版）》
CAS
1993年第1期45-49,共5页
Natural Science Journal of Hainan University
基金
海南省卫生厅科研基金
关键词
Β地中海贫血
基因分析
分子遗传病
β—Thalassemia, the qualitative osmostic fragilty test, the quatitation of HbA_2, Li nationality, polymerase chain reaction
