多重PCR用于缺失型α-地中海贫血的基因检测

Multiplex PCR for Detecting Genotypes of Deletional α-Thalassemia

为了探讨多重PCR技术在检测我国南方常见缺失型α 地中海贫血中的临床应用 ,观察缺失型α 地中海贫血的基因分布频率 ,采用单管多重DNA扩增的方法 (M PCR)对经血红蛋白定量分析初步诊断为标准型和静止型α 地中海贫血及血红蛋白H(HbH)病的 14 5例患者进行基因检测。扩增产物经 1.2 %琼脂糖凝胶电泳 ,出现 1.3及1.8kb条带者 ,提示为 SEA/αα ;1.6及 1.8kb条带者为 α4.2 /αα ;1.8及 2 .0kb条带者为 α3 .7/αα ;1.3及 1.6或 2 .0kb条带 ,提示为缺失型HbH病 ( SEA/ α4.2 或 SEA/ α3 .7)。结果表明 ,14 5例受检者中发现 10 0例 SEA/αα(6 8.9% ) ,15例 α3 .7/αα(10 .3% ) ,8例 α4.2 /αα(5 .5 2 % ) ,2例 α3 .7/ α4.2 (1.38% ) , α3 .7及 α4.2 纯合子各 1例 (0 .6 9% ) ;14例 SEA/ α3 .7(9 6 5 % ) ,2例 SEA/ α4.2 (1.38% ) ;另有 2例患者产前诊断证实为Bart水肿胎儿。结论 :运用多重PCR技术可以准确、简便、快速地检测我国南方地区常见的 α3 .7、 α4.2 、 SEA3种缺失型α 地中海贫血 ,这一技术对α 地中海贫血的大人群筛查及携带者的检出是一种较为理想的方法。

To investigate the clinical application of multiplex PCR in detecting genotypes of deletional α-thalassemia in South China and observe the distribution frequency of α-globin gene deletion,145 patients with silent carrier,alpha thalassemia trait or HbH were identified by M-PCR and 1.2% agarose gel electrophoresis. There are 1.3,1.6,1.8 and 2.0 kb bands which indicate -- SEA ,-α 4.2 ,αα and -α 3.7 ,respectively. The results showed that among 145 patients,100 patients with -- SEA /αα(68.9%),15 with -α 3.7 /αα(10.3%),8 with-α 4.2 /αα(5.52%),2 with -α 3.7 /-α 4.2 (1.38%),1 with-α 3.7 /-α 3.7 (0.69%),1 with -α 4.2 /-α 4.2 (0.69%),14 with-- SEA /-α 3.7 (9.65%),2 with -- SEA /-α 4.2 (1.38%)were found. Two patients prenatal diagnosed were confirmed with Bart′s hydrops fetuses. In conclusion,M-PCR analysis is a simple,rapid and accurate method for detection of α-thalassemia gene deletion. This technique is helpful in screening,carrier identification and prenatal diagnosis of deletional α-thalassemia.