荧光多重PCR对α地中海贫血的基因诊断

Diagnosis of several common α-thalassemia genotypes by multiplex fluorescence polymerase chain reaction

目的 :建立一种快速、灵敏的α地中海贫血诊断方法。方法 :应用荧光引物对正常人和患者进行多重PCR ,同时扩增α1和α2 珠蛋白基因和内对照β肌动蛋白 ( β actin) ,用PerkinElmerABIPRISMTM3 77DNASequencer自动分析结果 ,并对所得数据进行统计。用连锁分析的方法PCR扩增同一批模板α2 珠蛋白基因上游的 (CA) n,跑变性胶并将结果与前一种方法对照。结果 :荧光多重PCR自动分析和扩增 (CA) n 连锁分析得出的结论与实际情况完全相符 ,结果判断容易。结论 :荧光多重PCR法灵敏、简便 ,单管内就能完成 ,适合运用于α地中海贫血的快速诊断 ,并有可能运用于移植前诊断和母体外周血分离胎儿有核红细胞产前诊断。

Objective To develop an approach to a quick and accurate genetic diagnosis. Methods By using fluorescence primer,the fragments of α 1 and α 2-globin gene and internal control: β-actin were amplified and then the PCR products were analyzed with Perkin Elmer ABI PRISMTM 377DNA Sequencer.Linkage analysis was also performed with(CA) n at the upstream of α 2-globin gene. The two methods were compared. Result The fluoresence polymerase chain reaction and the linkage analysis gave the same result, and it was easy to distinguish different genotypes. Conclusion The multipilx fluoresence polymerase chain reaction is simple, sensitive and feasible for rapid diagnosis of α thalassemia,and may be utilized for the genetic diagnosis of the pre-transplantation and the diagnosis by fetal nucleated erythrocytes in maternal blood.