基于Nanopore的长读长测序对脆性X综合征一个家系的甲基化表观遗传学分析  

Methylation epigenetic analysis of a pedigree affected with Fragile X syndrome based on Nanopore long-read sequencing

作　　者：王聪慧 时盼来 刘莉娜 赵学潮 孔祥东[1] Wang Conghui;Shi Panlai;Liu Li′na;Zhao Xuechao;Kong Xiangdong(Genetics and Prenatal Diagnosis Center,the First Affiliated Hospital of Zhengzhou University,Zhengzhou,Henan 450052,China)

机构地区：[1]郑州大学第一附属医院遗传与产前诊断中心,郑州450052

出　　处：《中华医学遗传学杂志》2024年第11期1290-1295,共6页Chinese Journal of Medical Genetics

基　　金：国家重点研发计划(2018YFC1002206-2)。

摘　　要：目的应用Nanopore长读长测序技术探讨1个脆性X综合征(FXS)家系的遗传学病因。方法选取2023年4月于郑州大学第一附属医院进行遗传咨询的1个FXS家系为研究对象。应用Nanopore长读长测序、三核苷酸重复引物PCR(TP-PCR)、甲基化特异性多重连接探针扩增(MS-MLPA)及雄激素受体(AR)基因三核苷酸多态性基因型分析对家系中相应成员进行FMR1基因CGG重复数、甲基化和X染色体失活分析。本研究已通过郑州大学第一附属医院医学伦理委员会的审查(伦理号:KS-2018-KY-36)。结果先证者检出FMR1基因全突变且CpG岛甲基化;其胞姐检出1条X染色体FMR1基因全突变且CpG岛甲基化,另1条X染色体FMR1基因未见异常;其母亲检出FMR1基因前突变。结论基于Nanopore的长读长测序可同时检测女性2条X染色体的FMR1基因动态突变和甲基化情况,对不同性别FXS患者的诊断具有重要的临床价值。Objective To explore the genetic basis for a Chinese pedigree affected with Fragile X syndrome(FXS)through Nanopore long-read sequencing.Methods A FXS pedigree who had undergone genetic counseling at the First Affiliated Hospital of Zhengzhou University in April 2023 was selected as the study subject.Nanopore long-read sequencing,triplet-repeat primed PCR(TP-PCR),methylation-specific multiplex ligation-dependent probe amplification(MS-MLPA)and trinucleotide polymorphism genotyping of androgen receptor(AR)gene were used to analyze the FMR1 CGG repeat number,methylation,and X chromosome inactivation of the pedigree members.This study has been approved by the Medical Ethics Committee of the First Affiliated Hospital of Zhengzhou University(No.KS-2018-KY-36).Results Full mutation and CpG island hypermethylation were detected in the proband.The elder sister of the proband had full mutation of the FMR1 gene on one X chromosome and hypermethylation of CpG island,while the FMR1 gene on the other X chromosome was normal.FMR1 premutation was detected in the proband′s mother.Conclusion Nanopore long-read sequencing can simultaneously detect the dynamic mutation and methylation status of the FMR1 gene on the two X chromosomes of females,which has important value for the diagnosis of FXS in different genders.

关 键 词：长读长测序 脆性X综合征 FMR1基因 甲基化 动态突变 X染色体失活 

分 类 号：R596[医药卫生—内科学]