采用Amplicon文库构建法对地中海贫血的基因检测

Gene diagnostic of thalassemia by the method of constructing amplicon libraries

目的进一步完善现有地中海贫血的检测方法并且建立一种新的基于高通量测序的基因诊断方法,研究α/β地中海贫血基因的突变类型及分布情况。方法采用Amplicon文库构建法,基于高通量测序平台,对219例地中海贫血患者全血样本进行上机测序与数据分析。结果在219例地中海贫血患者中,共计检测出14种突变类型,包括CD122、CD125、CD142 3种非缺失型α-地贫突变及IVS-Ⅱ-654、CD41-42、CD17等11种常见β-地贫突变型别。此外,通过生物信息学软件作图分析,筛查出--^(SEA)、-α^(3.7)、-α^(4.2)3种缺失型α-地贫。结论本研究进一步完善了基于高通量测序的基因诊断方法,建立了一套Amplicon文库构建法,能准确、有效地用于诊断α/β地中海贫血基因的缺失与突变类型,对人群筛查,优生优育的遗传咨询,防止该病重症患儿的出生等具有重要意义。

Objective To further improve the existing diagnostic methods of thalassemia, establish a new method for gene diagnosis base on next generation sequencing and research the mutation type and distribution of alpha/beta thalassemia gene. Methods 219 cases of the whole blood samples with thalassemia have been sequenced and analyzed using the method of constructing amplicon libraries, which is based on next generation sequencing platform. Results There were total of 14 mutation types detected,including 3 non-deletion of CD122, CD125 and CD142 in alpha thalassemia, 11 kinds of common mutation of IVS-Ⅱ-654, CD41-42, CD17 and other in beta thalassemia in these 219 cases of the thalassemia patients. In addition, through graphing analysis by Bioinformatics software, 3 type of alpha deletion of--^SEA,-α^3.7 and-α^4.2 have been screened out. Conclusion This study have further improved the gene diagnostic method base on next generation sequencing technology and set up a method of constructing amplicon libraries that can be used for diagnosing the deletion and mutation types of alpha/beta thalassemia gene accurately and effectively. It has a great significance in population screening, genetic counseling of prenatal and postnatal care, disease preventing for newborn with severe mediterranean anemia, etc.