Geever R F, Wilson L B, Nallaseth F S, Milner P F, Bittner M, Wilson J T
Proc Natl Acad Sci U S A. 1981 Aug;78(8):5081-5. doi: 10.1073/pnas.78.8.5081.
Several reports have been published on the use of polymorphisms found in the human hemoglobin genes as a means for prenatal diagnosis of sickle cell anemia. The disadvantages of this approach reside in its limited application and the need for family analysis. Here we report that, by use of restriction endonuclease Dde I and diazobenzyloxymethyl-paper transfer procedures, a direct analysis can be made. Individuals with normal hemoglobin (AA) show two bands (175 and 201 base pairs) complementary to a 5'-specific beta-globin gene probe. Sickle cell trait individuals (AS) exhibit an additional band (376 base pairs). Individuals with sickle cell anemia (SS) show the band at 376 base pairs with a concomitant loss of the 175-base pair band. We interpret these changes in banding pattern to be the result of the elimination of a restriction site for Dde I in the altered codon associated with the sickle cell allele. Because an analysis can be performed on as little as 20 micrograms of cellular DNA, the application to prenatal diagnosis of sickle cell anemia should be possible.
已有多篇报道介绍了利用人类血红蛋白基因中的多态性进行镰状细胞贫血产前诊断的方法。这种方法的缺点在于其应用有限且需要进行家系分析。在此我们报告,通过使用限制性内切酶Dde I和重氮苄氧基甲基纸转移程序,可以进行直接分析。具有正常血红蛋白(AA)的个体显示出与5'-特异性β-珠蛋白基因探针互补的两条带(175和201个碱基对)。镰状细胞性状个体(AS)表现出一条额外的带(376个碱基对)。患有镰状细胞贫血(SS)的个体显示出376个碱基对的条带,同时175个碱基对的条带消失。我们将这些条带模式的变化解释为与镰状细胞等位基因相关的改变密码子中Dde I限制性位点消除的结果。由于只需对少至20微克的细胞DNA进行分析,因此有可能将其应用于镰状细胞贫血的产前诊断。