期刊论文

中文

临床检验杂志

1001-764X

2014

32

4

245-248

10.13602/j.cnki.jcls.2014.04.02

国家自然科学基金（81102516） 南华大学博士启动基金（2012XQD18）.

本校为其他机构

目的 用高保真DNA聚合酶介导的PCR检测β地中海贫血基因热点突变.方法 选取已知中国人群β珠蛋白基因CD41-42、IVS-2nt654、TATAbox-28、CD17、CD71-72及CD26热点突变位点为检测靶点,分别设计3'末端与野生型基因位点或突变型基因位点完全配对的引物,并用硫化磷酸修饰,进行高保真DNA聚合酶介导的引物延伸反应.用多重PCR反应体系检测临床已知分别含CD26和TATAbox-28突变热点患者的DNA标本.结果 对野生模板而言,仅有野生型等位基因相关引物有产物产生,而突变型等位基因位点特异性引物无产物产生.反之,使用突变模板,仅突变型等位基因位点相关引物有产物产生,而野生型等位基因位点特异性引物无产物产...

Objective To detect the hotspot mutations of 13-thalassemia by high fidelity DNA polymerase-mediated PCR. Methods CIM1-42, IVS-2nt654, TATAbox-28, CD17, CD71-72 and CD26 hotspot mutational sites in β-globin gene were selected as detection targets. Then, allelic specific primers for these wild or mutational alleles were designed and modified with phosphorothioate. Next, high fidelity DNA polymerase-mediated primer extension reaction was carried out. Moreover, the DNA samples containing CD26 or TATAhox-28 mutations from the patients with β-thal...