Dianzani I, Camaschella C, Saglio G, Forrest S M, Ramus S, Cotton R G
Istituto di Clinica Pediatrica, Torino, Italy.
Genomics. 1991 Sep;11(1):48-53. doi: 10.1016/0888-7543(91)90100-s.
We used the chemical cleavage of mismatch (CCM) method to screen the beta-globin gene simultaneously for Mediterranean beta-thalassemia mutations. The beta-globin gene was amplified in two segments encompassing the whole gene and hybridized to a corresponding labeled PCR product from a normal subject. All the known mutations tested were identified and discriminated. Three beta-thalassemic subjects with previously undiagnosed mutations were identified as carriers of two rare DNA changes. The inheritance of the mutations could be traced in family studies, showing the reliability of the method even for prenatal diagnosis. The beta-globin gene polymorphisms were also detected and the framework was determined for most alleles. Our results suggest further applicability of the CCM method as a means to screen a gene simultaneously for multiple mutations.
我们采用错配化学切割（CCM）方法同时筛查β-珠蛋白基因，以检测地中海β-地中海贫血突变。β-珠蛋白基因被扩增为两个片段，覆盖整个基因，并与来自正常受试者的相应标记PCR产物杂交。所有检测的已知突变均被识别和区分。三名先前未诊断出突变的β-地中海贫血受试者被鉴定为两种罕见DNA变化的携带者。在家族研究中可以追踪突变的遗传情况，这表明该方法即使用于产前诊断也具有可靠性。还检测到了β-珠蛋白基因多态性，并确定了大多数等位基因的框架。我们的结果表明CCM方法作为一种同时筛查基因多种突变的手段具有进一步的适用性。
