摘要
目的探讨高分辨熔解曲线(HRM)分析技术检测β-地中海贫血基因突变的可行性。方法应用HRM法检测广东省常见的5种β-地中海贫血突变(-28、CD17、CD41-42、CD71-72及IVS-2-654),对60例可疑β-地中海贫血患者样本进行HRM分析,通过DNA测序对HRM结果进行验证。结果 60例可疑β-地中海贫血患者中,共检出杂合突变型基因12例,包括2例-28、2例CD17、4例CD41-42、2例CD71-72及2例IVS-2-654基因突变类型;检出纯合突变型基因2例,包括1例-28及1例CD41-42(均为羊水标本)。HRM法检测结果与测序结果一致。结论 HRM法能准确检测广东省5种常见的β-地中海贫血突变,具有简便、快速、灵敏度高等优点,有望成为临床筛查β-地中海贫血突变的新方法。
Objective To investigate the feasibility of high resolution melting（HRM）curve analysis technique for detectingβ-thalassemia gene mutations.Methods The HRM method was used to detect five kinds of commonβ-thalassemia mutations（-28,CD17,CD41-42,CD71-72 and IVS-2-654）in Guangdong province.Sixty specimens of the patients with suspectedβ-thalassemia were performed the HRM analysis.The results of HRM analysis were confirmed by the direct DNA sequencing.Results Among 60 specimens of the patients with suspectedβ-thalassemia,12 cases of heterozygosis mutant gene were detected,including 3cases of-28,2cases of CD17,5cases of CD41-42,2cases of CD71-72 and 2cases of IVS-2-654 gene mutation;2cases of homozygosis mutant gene were detected,including 1case of-28 and 1case of CD41-42 homozygous mutation（both were amniotic fluid specimens）.The results of HRM analysis were consistent with the DNA sequencing results.Conclusion The HRM method can accurately detect five kinds of commonβ-thalassemia mutations in Guangdong province,has the advantages of simpleness,rapidness and high sensitivity and is expected to be a new method for screeningβ-thalassemia mutation in clinic.
出处
《国际检验医学杂志》
CAS
2015年第5期650-651,654,共3页
International Journal of Laboratory Medicine
基金
番禺区科技计划资助项目(2014-Z03-68)
