Rapid genotyping of known mutations and polymorphisms in β-globin gene based on the DHPLC profile patterns of homoduplexes and heteroduplexes

被引：34

作者：

Li, Qiang ^{[1
,2
]}

Li, Li-Yan ^{[1
]}

Huang, Sheng-Wen ^{[1
]}

Li, Liang ^{[1
]}

Chen, Xiao-Wei ^{[1
]}

Zhou, Wan-Jun ^{[1
]}

Xu, Xiang-Min ^{[1
]}

机构：

[1] So Med Univ, Sch Basic Med Sci, Dept Med Genet, Guangzhou 510515, Guangdong, Peoples R China

[2] So Med Univ, Nanfang Hosp, Dept Lab Med, Guangzhou 510515, Guangdong, Peoples R China

来源：

CLINICAL BIOCHEMISTRY | 2008年 / 41卷 / 09期

关键词：

denaturing high-performance liquid chromatography (DHPLC); beta-thalassemia; genotyping; mutation analysis; polymorphism;

D O I：

10.1016/j.clinbiochem.2008.02.008

中图分类号：

R446 [实验室诊断]; R-33 [实验医学、医学实验];

学科分类号：

1001 ;

摘要：

Background: beta-thalassemia represents a great heterogeneity as over 200 mutations have been identified for the beta-globin gene responsible for this disease. A rapid genotyping test with high accuracy, selectivity, and reproducibility suitable for the determination of known mutations is needed for prenatal screening and post-natal diagnosis of this disease in clinical setting. Design and methods: We have performed the validation of a DHPLC assay for direct genotyping of known causative mutations in beta-globin gene using the chromatographic pattern-based strategy under partially-denaturing conditions. Results: DHPLC assay was established based on the analysis of 795 DNA samples from a group of various genotypes for the 20 mutations and 8 polymorphisms in beta-globin gene then validated on 319 tests in a blind study. The results obtained with this assay were in concordance with the results obtained by DNA sequence analysis. Conclusion: This simple method can meet the requirements of direct genotyping of known beta-thalassemia mutations and/or polymorphisms in the clinical setting for Chinese and in general as a model for other populations. (c) 2008 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

引用

页码：681 / 687

页数：7

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