地高辛标记DNA探针在α-地中海贫血基因诊断中的应用

APPLICATION OF DIGOXIGENIN LABELED DNA PROBE FOR THE DETECTION OF GENIE DELETION IN α-THALASSEMIAS

以地高辛标记DNA片段作探针,用点杂交法可检出0.1pg目标DNA。消化5～10μg人基因组DNA,经印迹转移后与相应标记探针杂交,可得清晰的α—珠蛋白基因限制酶切图谱,试用本法于α—地贫基因分析,31例受检者的结果为:14例无α—珠蛋白基因缺失,10例为α—地贫1杂合子。5例为α—地贫2杂合子,2例为α—地贫1与α—地贫2杂合子。

A nonradioactive digoxigenin labeled probe was used to detect the gene deletion of α-thalassemias. As little as 0.1 pg of target DNA could be detected by digoxigenin labeled probe in dot blot hybridization. When southern blot hybridization was used, the α-globin gene maps could be dearly visualized form 5-10 μg of genomic DNA digested with appropriate restriction enzymes. The results of 31 cases examined by this method revealed that: in 14 cases no α-globin gene deletion detected, 10 cases were heterozygotes of α-thalassemia-1, 5 cases were heterozygotes of α-thalassemia-2, and 2 cases were heterozygotes of α-thalassemia-1 and α-thalassemia-2.