多重PCR检测缺失型α地中海贫血

Detection of deletional α-thalassemia by multiple PCR

目的 针对中国人常见的缺失型α地中海贫血建立一种快速、简便、可靠的多重聚合酶链反应 (PCR)基因诊断技术。方法 设计 4对引物 ,利用单管多重PCR方法进行扩增 ,同时检测 3种缺失型和正常的α2 珠蛋白基因。结果 成功检测出正常人及 3种缺失型的杂合子、双重杂合子 ;正常α2 基因的扩增片段为 180 0bp ,东南亚型缺失基因的扩增片段为 70 8bp ,右侧缺失基因的扩增片段为 2 0 2 2bp ,左侧缺失基因的扩增片段为 16 2 8bp , α3 .7/ SEA双重杂合子的扩增片段为 2 0 2 2bp和 70 8bp , α4.2 / SEA双重杂合子的扩增片段为 16 2 8bp和 70 8bp。结论 790例标本的基因检测结果分析表明 ,该法快速、准确 ,可用于常规临床标本的检测。

Objective To establish a rapid,simple and reliable multiplex-polymerase chain reaction(PCR) method for detection of deletional α-thalassemia in Chinese population.Methods Four pairs of primers were designed and multiplex-PCR was amplified in a single-tube to detect the genes of α 2 globin in three types of deletional α-thalassemia as well as normal α 2 globingene.Results The genes of normal α 2 globin and in deletional α-thalassemia were detected,and 3 types of heterozygote and double heterozygotewere were successfully identified.The fragment of normal α 2 globin gene was 1 800 bp;while the fragment of SEA (southeastern asia)-type of deletional thalassemia gene was 708 bp,the rightward deletion [-alpha(3.7)] fragment was 2 022 bp,and the leftward deletion[-alpha(4.2)] fragmentwas 1 628 bp.The fragments of SEA-type/-alpha(3.7) compound heterozygotes were 708 bp and 2 022 bp,and the fragments of SEA-type/-alpha(4.2) compound heterozygotes were 708 bp and 1 628 bp.Conclusions The results of analysis for 790 samples proved the multiplex-PCR is a rapid,simple and reliable method for detection of alpha-thalassemia in clinical laboratory.