- 作者: 李賢雄; 張建國; 陳龍德; 楊勉力; 朱廣邦
- 作者服務機構: 台北榮民總醫院醫學研究部基因重組室; 台北榮民總醫院婦產部; 台北新立仁愛醫院分子醫學暨臨床病理部及病理中心
- 中文摘要:
本研究是結合多片段單股核酸多元性結構性(MRF-SSCP)及擴增衍生限制?切割點(ACRS)兩種技術,應用於偵測
國人常見的β型地中海型貧血症的產前診斷。前者的方法,是使用一對引子(primor)及帶有核標誌的核酸?,擴增β球
蛋白基因。隨後以三種限制?(BbSI, BamHI,及DraI)切割。所得的片段,在含有甘油之聚丙烯醯胺電泳膠體分析。其
結果與利用擴增衍生限制?切割點一致。同時,以單股核酸多元性結構型分析及利用單向擴增核酸定序(LADS)發現,
二個國人常見的多元性位置。其中,在密碼-3(CAT或CAC)是一未曾報告的鹼基改變。此二種基因點改變與突變型
位置的交互出現,對於利用單股核酸多元性的分析產生影響。 - 英文摘要: A modified approach of polymerase chain reaction (PCR)-based single strand conformation poly-morphism (SSCP) has been developed for prenatal diagnosis of β-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 SSCPgel. The method is thus termed multiple restriction fragment (MRF)-SSCP. Two cases of chorionic samplesand the blood samples of their parents were studied. The previously described amplification-created re-striction site (ACRS) method was used to confirm the SSCP results obtained. Two polymorphic sites ofthe β-globin gene which would influence the MRF-SSCP patterns are described, including a previouslyundescribed polymorphic site located at codon 3 (CAT or CAC) of the β-globin gene.
- 中文關鍵字: β-thalassemia; single strand conformation polymorphism; prenatal diagnosis.
- 英文關鍵字: --