四川乐山地区地中海贫血基因突变类型分析研究  

Analysis of thalassemia gene mutation types in Leshan Sichuan

作　　者：陈丽龙 CHEN Li-long(Department of Laboratory,Leshan People's Hospital,Sichuan Leshan 614000)

机构地区：[1]四川乐山市人民医院检验科,四川乐山614000

出　　处：《医学检验与临床》2020年第5期27-30,共4页Medical Laboratory Science and Clinics

摘　　要：目的:通过调查四川乐山地区人群中α和β地中海贫血(简称地贫)的发生率、基因缺陷类型及构成比,为该地区开展地贫基因产前诊断,预防重型地贫患儿出生及优生优育事业的发展奠定基础。方法:通过收集2018年1月~2019年12月在我院检验科分子实验室进行产前地贫基因检测的病例共4406例,应用聚合酶链反应(polymerase chainreaction,PCR)结合反向斑点杂交技术(reverse dot blot,RDB),通过制备DNA标本,进行PCR扩增、杂交、洗膜等步骤,最后显色观察结果,分析地贫基因突变类型,对基因型频率分布进行归纳总结。结果:在4406份外周血标本中,共检出26种基因型,阳性检出率为13.78%(607例)。其中,检出缺失型α地贫330例,其中SEA/αα(128例),-α3.7/αα(175例),-α4.2/αα(15例),构成比分别为38.79%、53.03%、4.5%,基因频率最高的是-α3.7/αα。检出非缺失型α地贫16例,其中CD122(CAC-CAG)5例,CD142(TAA-CAA7)7例,CD125(CTG-CCG)4例,共占2.6%。检出突变型β地贫2527例,共计发现基因型13种,其中最常见的突变类型分别为CD41/42(-TCTT)、CD17(A→T)、IVS-Ⅱ-654(C→T)、-28(A→G)、CD71/72(+A)、βE、共占41.51%,其中以CD17(A→T)位点突变所占比例最高,占17.46%,IVS-Ⅱ-654(C→T)次之,占11.77%,α地贫兼β地贫9例,占1.5%7,双重缺失型α地贫10例,基因型为-SEA/-α3.77,α纯合子2例,-α3.7/-α3.7。结论:①乐山地区地中海贫血的基因突变类型呈多样化,α地贫以-α3.7/αα最常见,β地贫以CD17(A→T)最常见②α地贫发病率高于β地贫。③RDB-PCR技术在地贫的检测中特异性及敏感性高,适合于中国人群常见的地贫基因检测。本研究可为本地区开展地中海贫血诊断和研究提供参考资料。Objective:To investigate the incidence,type and composition ratio ofαandβthalassemia(thalassaemia)in the population of Leshan,Sichuan,we carried out prenatal diagnosis of thalassemia genes in this area,and prevented the birth and eugenic birth of children with severe thalassemia Laid the foundation for development.Methods:A total of 4406 cases of prenatal thalassaemia gene testing in the molecular laboratory from January 2018 to December 2019 were collected.Polymerase chain reaction(PCR)combined with reverse dot blot hybridization was used.Reverse dot blot(RDB)technology,through the preparation of DNA samples,PCR amplification,hybridization,membrane washing and other steps,and finally color observation results,analysis of thalassemia gene mutation types,and summarize the genotype frequency distribution.Results:A total of 26 genotypes were detected in 4406 peripheral blood samples,and the positive detection rate was 13.78%(607 cases).Among them,330 cases of missing alpha thalassemia were detected,SEA/αα(128 cases),-α3.7/αα(175 cases),-α4.2/αα(15 cases),and the composition ratios were 38.79%,53.03%and 4.5%the highest gene frequency was-α3.7/αα.16 non-deleted alpha thalassaemias were detected,of which 5 were CD122(CAC-CAG),7 were CD142(TAA-CAA),and 4 were CD125(CTG-CCG)accounted for 2.6%.252 cases of mutant beta thalassemia were detected,and a total of 13 genotypes were found.The most common mutation types were CD41/42(-TCTT),CD17(A→T),and IVS-II-654(C→T),-28(A→G),CD71/72(+A),βE,accounted for 41.51%,CD17(A→T)site mutation accounted for the highest proportion 17.46%,followed by IVS-II-654(C→T)accounted for 11.7%,9 cases of alpha thalassemia and beta thalassemia accounted for 1.5%,10cases of double deletion alpha thalassaemia,genotype-SEA/-α3.7.Conclusion:①The types of gene mutations in thalassaemia in Leshan area are diversified,α-thalassemia is most common with-α3.7/αα,andβ-thalassemia is most common with CD17(A→T).②α-thalassemia has higher incidence than Beta thalassaemia.③RDB

关 键 词：地中海贫血 基因突变 反向斑点杂交法 四川乐山 

分 类 号：R55[医药卫生—血液循环系统疾病]