摘要
为探讨临床对β地中海贫血（β－地贫）的快速基因诊断方法，应用ＰＣＲ反向点杂交（ｒｅｖｅｒｓｅｄｏｔｂｌｏｔ，ＲＤＢ）技术，对广东地区３５例重型β地贫患儿及其双亲的基因突变特征进行了研究。结果显示：（１）广东重型β地贫基因突变可见７种类型：ＣＤ４１－４２（－ＴＣＴＴ），ＩＶＳ２ｎｔ６５４（Ｃ→Ｔ）。ＴＡＴＡｂｏｘ－２８（Ａ→Ｇ），ＣＤ１７（Ａ→Ｔ），βＥ（２６）（Ｇ→Ａ），ＣＤ７１－７２（＋Ａ）及ＣＤ１４－１５（＋Ｇ），其结构比依次为：４０％，２８．６％，１１．４％，７．１％，７．１％，７．１％，４．３％及１．４％，和１３种基因组合形式；（２）基因突变类型不同致临床表型不同，β°纯合子临床多表现重症，发病早（３～６个月），靠输血维持生命；β°／β－（Ｅ或－２８）双重杂合子多表现中间型或重型，发病年龄较晚，贫血较轻，输血较少，ＰＣＲ－ＲＤＢ技术不需依赖同位素、操作简便、快速，宜于临床推广应用。
We have studied the molecular characterization of β-globin gene mutations in 35 children with β-thalassemia major and their parents in south China with the method of polymerase chain reaction(PCR) and reverse dot blot(RDB).The latter includes a set of allele specific digonucleotide(ASO) probes which can detect all 18 β-thalassemia mutations found in Chinese, These probes were immobilized on two strips of membrane, which were hybridized with β-globin gene sequences amplified by PCR with biotinlated primers and then treated with streptavidin-HRP and substrates for color development. The results indicated:(1) seven different mutations were identified among the patients with β-thalassemia in south China, namely CD 41-42(-TCTT). IVS2nt-654(C→T), TATA box-28(A→G),CD17(A→T),βE26(G→A),CD71-72(+A)and CD14-15(+G), with the frequencies of 40%, 28.6%.11.4%,7.1%, 7.1%, 4.3% and 1.4% respectively; (2)The dual heterozygens βE26(G→A) β-thalassemia patients had a milder clinical course;(3) The dual heterozygens -28 TATA box mutation β-thalassemia patients had a variation from severe clinical course to moderate one; (4)PCR-RDB is independent of radioisotopes, simple and rapid; it may be used as a routine method in clinical laboratories.
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
1996年第5期262-265,共4页
Chinese Journal of Medical Genetics
