高通量测序技术在新生儿地中海贫血基因筛查中的应用  

Application of Next Generation Sequencing to Screen the Neonatal Thalassemia Genes

作　　者：谭梅[1] 卢森 吴柳松[1] 靳大卫 彭智宇 陈艳[1] 

机构地区：[1]遵义医学院附属医院儿内二科,贵州遵义563000 [2]深圳华大基因,广东深圳518000

出　　处：《中国实验血液学杂志》2015年第5期1404-1409,共6页Journal of Experimental Hematology

基　　金：贵州省联合重点项目(黔科合J字LNK-2013-15)

摘　　要：目的:探讨高通量测序(NGS)技术在新生儿地中海贫血基因筛查中的可行性。方法:随机采集我院出生的206例新生儿足底血,制成干血片,用高通量测序(NGS)技术进行地中海贫血基因筛查,根据其结果进行进一步的分析。结果:206例受检新生儿中筛查出地中海贫血基因突变的有22例,其中α-地中海贫血11例,β-地中海贫血11例,新发5例。11例α-地中海贫血以基因缺失为主,共7例,所占比例达到64%(7/11),具体基因型分布:右侧缺失型(αα/-α3.7)4例,东南亚型(αα/-SEA)2例,左侧缺失型(αα/-α4.2)1例,其余4例为基因点突变所致,占36%(4/11),分别为:Hb Part-Dieu杂合、Hb Quong Sze杂合、Hb Westmead杂合、HBA1:c.95+9C>T(新发突变);11例β-地中海贫血主要为β-链基因点突变所致,共检出7种突变基因型;以CD17(A→T)位点的突变最多见,占27%(3/11),-50 G>A杂合2例、Hb Hamilton杂合1例、IVS-II-654(C→T)1例,其余4种为新的基因位点突变,分别为:HBB:c.316-116C>A、HBB:c.316-248G>T、HBB:c.315+63T>C、HBB:c.-23 A>G。结论:高通量测序技术对新生儿足底干血片进行地中海贫血基因筛查是可行的,不仅能有效地检出地中海贫血的基因类型,而且还能发现新的基因突变。本方法具有采集标本简单、结果精确度高等优点,可广泛应用于临床,为地中海贫血的早期诊断提供保障。Objective: To explore the feasibility of using next-generation sequencing technology( NGS) to screen the neonatal thalassemia genes. Methods: Plantar blood of 206 cases of neonatal born in our hospital were randomly collected to be made into dried blood,which can be screened for thalassemia genes by next-generation sequencing,and then a further analysis would be performed on the basis on the detection results. Results: In 206 cases of neonates tested,the thalassemia gene mutations in 22 cases were screened,including 11 cases of alpha-thalassemia,11 cases of beta-thalassemia,5 cases of new mutations. Out of 11 cases of alpha-thalassemia 7 cases were proved to be the gene deletion,accounting for 64%( 7 /11),and the specific genotype distribution was as follows: 4 cases of αα/- α3. 7,2cases of αα/- SEA,1 case of αα/- α4. 2,the remaining 4 cases with point mutations( 4 /11,36%) : Hb Part-Dieu hybrid,Hb Quong Sze hybrid,Hb Westmead hybrid,HBA1: c. 95 + 9 c > T( rewly discovered gene mutation).The whole 11 cases of β-thalassemia are proved to be with beta chain point mutations,7 kinds of mutation genotype were detected,CD17( A- > T) is the most common point locus mutation,accounted for 27%( 3 /11),and 50 G > A hybrid in 2 cases,1 cases of Hb Hamilton hybrid,IVS- II- 654( C- > T) in 1 case. The remaining 4 cases are of the new gene point mutation,they are as follows respectively: HBB: c. 316- 116 c > A,HBB: c. 316- 248 G > T,HBB:c. 315 + 63 T > c,HBB: c.- 23 A > G. Conclusion: The next-generation sequencing technology can be used to screen neonatal plantar dried blood for the thalassemia genetic mutation,which not only can effectively detect thalassemia gene types,but also can look for new gene mutations. The advantages of this method include easy collecting samples,precise result and wide use for clinical diagnosis,thus possibly give an early diagnosis for thalassemia.

关 键 词：地中海贫血 高通量测序技术 新生儿足底干血片 基因筛查 

分 类 号：R722.1[医药卫生—儿科]