变性高效液相色谱法检测中国南方和东南亚人群常见缺失与突变型α-地中海贫血  

Detection of common deletions and mutations causing α-thalassemia in Southeast Asians and Southern Chinese with denaturing high performance liquid chromatography

作　　者：贾兴元[1,2] 卫小静[1,3] 唐宁 王立荣[2] 韩瀚[1] 郑梅玲[3] 蔡稔 肖白[1] 刘敬忠[1] 

机构地区：[1]首都医科大学附属北京朝阳医院实验研究中心,北京100020 [2]中国医学科学院＆北京协和医学院基础医学院医学遗传室 [3]山西医科大学附属第一医院计划生育研究室 [4]广西柳州市妇幼保健院遗传优生科

出　　处：《中华医学遗传学杂志》2011年第6期670-674,共5页Chinese Journal of Medical Genetics

基　　金：基金项目：北京市自然科学基金（7092034）

摘　　要：目的针对中国南方和东南亚人群α-地中海贫血常见缺失与点突变类型，建立一种简便、准确与快速的两管PCR结合变性高效液相色谱检测方法。方法双重聚合酶链反应（polymerasechainreaction，PCR）结合变性高效液相色谱（denaturinghighperformanceliquidchromatography，DHPLC）分析，联合改进型一管PCR结合DHPLC方法，对160份临床标本中α珠蛋白基因的3种常见点突变与3种缺失进行检测。结果用一管双重PCR—DHPLC和改进型一管PCR—DHPLC分析法分别检测了α珠蛋白基因常见的3种点突变与3种缺失，检测结果与临床现行方法相符。结论改进的检测方法可同时检测中国南方和东南亚人群α-地中海贫血的常见缺失与点突变，具有准确、快速、半自动化及经济等特点，适用于规模化的临床筛查与检测。Objective To establish a comprehensive and simple assay using denaturing high performance liquid chromatography （DHPLC） for the diagnosis of most common mutations and deletions of α-thalassemia gene in Southeast Asians and Southern Chinese. Methods This assay has included a duplex polymerase chain reaction （PCR） followed by DHPLC analysis. An improved PCR was also performed followed by DHPLC analysis. With this assay, a blinded study of 160 samples was screened for three common mutations and three common deletions. Results The duplex PCR-DHPLC combined with the improved PCR-DHPLC analysis has detected all mutations and the wild-type allele. The results were consistent with those by the original methods. Conclusion This molecular assay may be used for the diagnosis of α-thalassemia patients from this geographical region. The method is accurate, rapid, semiautomatic and cost-effective, which makes it suitable for large-scale screening.

关 键 词：Α-地中海贫血 变性高效液相色谱 聚合酶链反应 分子诊断 

分 类 号：R440[医药卫生—诊断学]