Lee H H, Chang J G, Chen R T, Yang M L, Choo K B
Department of Medical Research, Veterans General Hospital-Taipei, Taiwan, Republic of China.
Proc Natl Sci Counc Repub China B. 1994 Jul;18(3):112-7.
A modified approach of polymerase chain reaction (PCR)-based single strand conformation polymorphism (SSCP) has been developed for prenatal diagnosis of beta-thalassemic mutations. In this method, a single PCR product (1,350-bp) is first generated, which is then digested by restriction enzymes (BbsI, BamHI, DraI) to generate multiple shorter restriction fragments for electrophoretic analysis in an SSCP gel. The method is thus termed multiple restriction fragment (MRF)-SSCP. Two cases of chorionic samples and the blood samples of their parents were studied. The previously described amplification-created restriction site (ACRS) method was used to confirm the SSCP results obtained. Two polymorphic sites of the beta-globin gene which would influence the MRF-SSCP patterns are described, including a previously undescribed polymorphic site located at codon 3 (CAT or CAC) of the beta-globin gene.
已开发出一种基于聚合酶链反应（PCR）的单链构象多态性（SSCP）改良方法用于β地中海贫血突变的产前诊断。在该方法中，首先生成单一的PCR产物（1350bp），然后用限制性内切酶（BbsI、BamHI、DraI）进行消化，以产生多个较短的限制性片段，用于在SSCP凝胶中进行电泳分析。因此，该方法被称为多限制性片段（MRF）-SSCP。研究了两例绒毛膜样本及其父母的血液样本。使用先前描述的扩增产生限制性位点（ACRS）方法来确认所获得的SSCP结果。描述了两个会影响MRF-SSCP模式的β珠蛋白基因多态性位点，包括一个位于β珠蛋白基因密码子3（CAT或CAC）处先前未描述的多态性位点。
