Bionano-DNA全长光学图谱技术在血友病A携带者基因诊断中的应用  

Application of Bionano-DNA Full-Length Optical Mapping Technology in Genetic Diagnosis of Hemophilia A Carriers

作　　者：牛思颖 刘梦艳[2] 丁子轩[1] 解珺丹[1] 江艾蕊 余自强[1] 王兆钺[1] 阮长耿[1] 夏利军[1] 江淼[1,3] NIU Si-Ying;LIU Meng-Yan;DING Zi-Xuan;XIE Jun-Dan;JIANG Ai-Rui;YU Zi-Qiang;WANG Zhao-Yue;RUAN Chang-Geng;XIA Li-Jun;JIANG Miao(Jiangsu Institute of Hematology,The First Affiliated Hospital of Soochow University,Suzhou 215007,Jiangsu Province,China;Deparment of Hematology,The First filiated Hospital of Soochow University,Suzhou 215007,Jiangsu Province,China;DushuLake Hospital filiated 10 Soochow University,Suzhou 215021,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院江苏省血液研究所,江苏苏州215007 [2]苏州大学附属第一医院血液科,江苏苏州215007 [3]苏州大学附属独墅湖医院,江苏苏州215021

出　　处：《中国实验血液学杂志》2023年第4期1211-1216,共6页Journal of Experimental Hematology

基　　金：江苏省自然科学基金项目(BK20201170);江苏省高校自然科学基金重点项目(20KJA320001);苏州市科技发展计划项目(SZM2021001)。

摘　　要：目的:尝试将Bionano Saphyr可视化全长DNA光学图谱技术应用于血友病A携带者的精准基因诊断。方法:针对2例应用常规二代测序技术无法诊断的可疑F8基因缺陷女性携带者,采用DNA全长光学图谱技术,检测扫描获得样本X染色体全长可视化单倍型特征性图谱,与正常人群DNA图谱库进行比对,获得待测样本的基因结构变异信息。结果:2例检测样本F8基因的X染色体DNA分子平均荧光标记长度大于2.5 Mbp,平均拷贝数大于20×。经比对分析,其中1例样本为F8基因22号内含子近端倒位,1例为22号内含子倒位同时伴有多处大片段缺失。结论:Bionano技术对于长度大、变异复杂的基因缺陷有很好的检出率。在无先证者或缺少先证者精准基因诊断结果的情况下,应用该技术检出F8基因的杂合复杂变异对血友病携带者的优生优育、孕前诊断有重要意义。Objective:To apply Bionano Saphyr visual fll-ength DNA optical mapping technology to the precise genetic diagnosis of hemophilia A carriers.Methods:For 2 suspected F8 gene deficiency female cariers who could not be diagnosed by conventional next-generation sequencing technology,the full-length DNA optical mapping technology was used to detect and scan the sample X chromosome full-length visual haplotype characteristic map,which was compared with the nomal haplotype.The gene structure variation information of the samples was obtained by compare with DNA atlas library.Results:The average fluorescent marker length of the X chromosome DNA molecular where the F8 gene was located in the two samples was greater than 2.5 Mbp,and the average copy number was greater than 20 x.After comparative analysis,one of the samples was a proximal inversion of intron22 of the F8 gene,and another was an inversion of intron 22 accompanied by muliple deletions of large fragments.Conclusions:Bionano technology has a good detection rate for gene defects with large length and complex variation.In the absence of a proband or accurate genetic diagnosis results of the proband,the application of this technology to detect the heterozygous complex variant of the F8 gene is of great significance for the prenatal diagnosis and pre-pregnancy diagnosis of hemophilia carriers.

关 键 词：Bionano技术 血友病A 内含子倒位 基因诊断 

分 类 号：R554.1[医药卫生—血液循环系统疾病]