应用反向点杂交法检测α-地中海贫血点突变  

Detection of the Point Mutations of α-Thalassemia by Reverse Dot Blot

作　　者：郭广洲[1] 陈延娥[2] 廖生赟 周敏 

机构地区：[1]深圳市第五人民医院检验科,深圳518001 [2]北京大学深圳医院检验科,深圳518034 [3]亚能生物技术(深圳)有限公司,深圳518034

出　　处：《热带医学杂志》2008年第8期785-787,共3页Journal of Tropical Medicine

基　　金：深圳市科技局课题(No.2005-196)

摘　　要：目的建立一种临床上切实可行的非缺失型α-地中海贫血基因检测方法。方法中国人常见的非缺失α-地中海贫血为αCSα、αQSα、αWSα3种类型,针对此3个位点设计特异性带有生物素标记的引物进行PCR扩增,扩增产物与固定在膜条上的寡核苷酸探针进行杂交,并通过一系列显色反应分析结果。结果分别分析3种突变类型的正常及突变位点信号,明确检测样品在3个位点是否发生了突变,及是否为突变纯合子或杂合子,验证了该方法可诊断非缺失型!-地中海贫血点突变。结论该检测方法结果可靠,且简便易行,对实验室仪器设备要求不高,适合于在临床推广。Objective To establish a convenient and reliable method to detect nondeletional point mutations of ct-thalassemia for clinical diagnosis. Method Genotypes of αCSα, αQSα,αWSα polymorphisms of α-globulin gene are the most common types of nondeletional α-thalassemia in China. Biotin labeled primers specific to ctCSct, αCSα and ctWSct mutations were designed to amplify the fragment of α-globulin gene. A set of oligo nucleotide probes were immobilized on the nylon membrance to capture biotin labeled amplification products by hybridization. Color reactions were followed to obtain the final results. Result The colorful signals of wildtype or mutants could be observed by eyes easily.The existence and absence of the signals specific to three genotypes could indicate homozygote or heterozygote. This method to detect nondeletional point mutatious of α-thalassemia be tested. Conclusion Reverse Dot Blot was a reliable and convenient and easily applied detection system in clinical laboratories without complex instructments. It could be used as a common method in rapid typing of nondeletional α-thalassemia point mutations in China.

关 键 词：Α-地中海贫血 点突变 反向点杂交 

分 类 号：R556.61[医药卫生—血液循环系统疾病]