基于孕妇血浆cffDNA高通量测序对巴氏胎儿水肿综合征进行产前检测的研究  

Prenatal examination on Hb Bart’s hydrops fetalis syndrome with cffDNA from maternal plasma by high-throughput sequencing

作　　者：陈宏健 周俏苗 王游声[3] 李雅璇 符免艾 CHEN Hongjian;ZHOU Qiaomiao;WANG Yousheng;LI Yaxuan;FU Mian’ai(Department of Medical Genetics and Prenatal Diagnosis,Haikou Maternal and Child Health Hospital,Haikou,Hainan Province,570203;Department of Medical Genetics and Prenatal Diagnosis,Hainan Women and Children’s Medical Center,Haikou,Hainan Province,570000;Medical Genetics Centre,Guangdong Women and Children Hospital,Guangzhou,Guangdong Province,511400;Department of Reproductive Medicine,First Affiliated Hospital of Hainan Medical University,Haikou,Hainan Province,570102,China;Center of Reproductive Medicine,Haikou Maternal and Child Health Hospital,Haikou,Hainan Province,570203)

机构地区：[1]海口市妇幼保健院医学遗传与产前诊断科,海口570203 [2]海南省妇女儿童医学中心医学遗传与产前诊断科,海口570000 [3]广东省妇幼保健院医学遗传中心,广州511400 [4]海南医学院第一附属医院生殖医学科,海口570102 [5]海口市妇幼保健院生殖医学中心,海口570203

出　　处：《陆军军医大学学报》2023年第24期2563-2569,共7页Journal of Army Medical University

基　　金：海南省科技专项(ZDYF2020113);海口市科技计划(2022-033);海南省高等学校科学研究项目(Hnky2019-41)。

摘　　要：目的通过对孕妇血浆中胎儿游离DNA(cell-free fetal DNA,cffDNA)的深度测序实现无创性α-地中海贫血(--SEA型)的基因分型。方法对cffDNA进行α-地中海贫血基因分型。于2021年11月至2022年11月通过分离来自海口市妇幼保健院、海南省妇女儿童医学中心、海南医学院第一附属医院因夫妻双方为东南亚型α-地中海贫血(--^(SEA)/αα)而需要进行产前诊断的34例孕妇(孕12~24周,均为单胎妊娠)血浆,历经cffDNA的提取、文库构建与杂交、模板制备与富集、上机测序等一系列基于Ion Proton平台的高通量测序操作,采用多维贝叶斯概率模型方法评估胎儿患巴氏胎儿水肿综合征的概率。随后从孕妇羊水中提取DNA,利用Gap-PCR对α-地中海贫血进行基因分型,并比较cffDNA与胎儿羊水DNAα-地中海贫血基因分型的符合率。结果34例孕妇血浆的cffDNA分型结果与对应的羊水DNA分型结果进行比较,28例的分型结果一致,4例不一致,2例因孕妇未行产前诊断无法比较,最终结果相符率为87.5%。结论利用孕妇血浆cffDNA进行巴氏胎儿水肿综合征(--^(SEA)/--^(SEA))的鉴定获得成功,其与羊水DNA分型结果具有高水平的符合率。ObjectiveTo perform the noninvasive genetic typing ofα-thalassemia(--^(SEA))by deep sequencing of cell-free fetal DNA(cffDNA)in maternal plasma using high-throughput sequencing technology.Methodsα-thalassemia genotyping was performed on cffDNA maternal plasma of 34 pregnant women was isolated from bloods.It was performed that a series of next generation sequencing operations based on Ion Proton platform,including cffDNA extraction,library construction and hybridization,template preparation and enrichment,and sequencing.Multivariate Bayesian probability model was used to evaluate the probability of Hb Bart’s hydrops fetalis syndrome of fetus.Subsequently,fetal DNA was extracted from amniotic fluids of pregnant women.Gap-PCR was performed to genotypeα-thalassemia,and the coincidence rate of cffDNA from maternal plasma was compared with that of the fetal DNA from amniotic fluids.ResultsThe results of cffDNA genotyping in plasma of 34 pregnant women were compared with those of amniotic fluid DNA.Twenty-eight cases were consistent,4 cases were inconsistent,and 2 cases were unknown because the pregnant women did not receive prenatal diagnosis.The ultimate coincidence rate was 87.5%.ConclusionThe identification of Hb Bart’s hydrops fetalis syndrome(--^(SEA)/--^(SEA))is successful with a high level of coincidence rate through cffDNA in plasma of pregnant women.

关 键 词：巴氏胎儿水肿综合征 胎儿游离DNA 高通量测序 

分 类 号：R446.11[医药卫生—诊断学] R591.1[医药卫生—临床医学] R714.55