错配特异性内切酶法与变性高效液相色谱法在鉴定HBB基因突变中的比较
Comparison of the mismatch-specific endonuclease method and denaturing high-performance liquid chromatography for the identification of HBB gene mutations.
作者信息
Hung Chia-Cheng, Su Yi-Ning, Lin Chia-Yun, Chang Yin-Fei, Chang Chien-Hui, Cheng Wen-Fang, Chen Chi-An, Lee Chien-Nan, Lin Win-Li
机构信息
1Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei, Taiwan.
出版信息
BMC Biotechnol. 2008 Aug 12;8:62. doi: 10.1186/1472-6750-8-62.
BACKGROUND
Beta-thalassemia is a common autosomal recessive hereditary disease in the Meditertanean, Asia and African areas. Over 600 mutations have been described in the beta-globin (HBB), of which more than 200 are associated with a beta-thalassemia phenotype.
RESULTS
We used two highly-specific mutation screening methods, mismatch-specific endonuclease and denaturing high-performance liquid chromatography, to identify mutations in the HBB gene. The sensitivity and specificity of these two methods were compared. We successfully distinguished mutations in the HBB gene by the mismatch-specific endonuclease method without need for further assay. This technique had 100% sensitivity and specificity for the study sample.
CONCLUSION
Compared to the DHPLC approach, the mismatch-specific endonuclease method allows mutational screening of a large number of samples because of its speed, sensitivity and adaptability to semi-automated systems. These findings demonstrate the feasibility of using the mismatch-specific endonuclease method as a tool for mutation screening.
背景
β地中海贫血是地中海地区、亚洲和非洲地区常见的常染色体隐性遗传性疾病。β珠蛋白（HBB）基因已发现600多种突变，其中200多种与β地中海贫血表型相关。
结果
我们使用两种高度特异性的突变筛查方法，错配特异性内切酶和变性高效液相色谱法，来鉴定HBB基因中的突变。比较了这两种方法的敏感性和特异性。我们通过错配特异性内切酶方法成功区分了HBB基因中的突变，无需进一步检测。该技术对研究样本的敏感性和特异性均为100%。
结论
与变性高效液相色谱法相比，错配特异性内切酶方法因其速度、敏感性以及对半自动化系统的适应性，能够对大量样本进行突变筛查。这些发现证明了使用错配特异性内切酶方法作为突变筛查工具的可行性。
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