基于母体血浆单体型分析无创产前检测脊髓性肌肉萎缩症的研究  

Haplotype-Based Noninvasive Prenatal Testing of Spinal Muscular Atrophy

作　　者：罗凯[1] 陈敏[1] 卢森 赖峥菲[1] 曾艳欣 陈超 袁媛 王垚燊 李世泉 高雅 陈芳 阿叁 陈敦金[1] 

机构地区：[1]广州医科大学附属第三医院广州妇产科研究所,广东广州510150 [2]深圳华大基因研究院,广东深圳518083 [3]天津华大基因研究院翻译组学中心,天津300308 [4]广州华大基因研究院,广东广州510006

出　　处：《实用妇产科杂志》2017年第9期674-678,共5页Journal of Practical Obstetrics and Gynecology

基　　金：国家自然科学基金(编号:81671470);广东省科技计划项目(编号:2013B022000005);广东省自然科学基金(编号:2016A030313610)

摘　　要：目的:探索无创产前检测(NIPT)脊髓性肌肉萎缩症(SMA)在临床应用的可行性。方法:招募生育过SMA先证者且再次妊娠的24个家系,其家系中夫妻双方及先证者致病突变经多重连接探针扩增技术(MLPA)测定。采集家系外周血及孕11周以后的孕妇血浆,通过目标序列捕获及高通量测序技术,首先对夫妻双方及先证者外周血DNA样品进行目标区域捕获测序,获得与致病位点连锁的亲本单体型信息,再结合血浆测序数据中提供的单核苷酸多态性位点的分析结果,构建隐马尔可夫模型,推断胎儿单体型,从而得到胎儿的基因型。同时利用有创产前诊断MLPA检测验证其准确性。本次研究主要针对SMN1基因第7、8号外显子的检测。结果:24个家系通过NIPT检测检出患胎5例,携带者7例,余12例正常;MLPA检测结果为5例患胎,8例携带者,11例正常。患胎均为SMN1基因第7、8号外显子纯合缺失。NIPT与MLPA检测结果一致率为95.83%(23/24),两种方法诊断结果差异无统计学意义(P=0.317)。结论:基于母体血浆目标区域捕获测序、单体型分析NIPT胎儿SMA风险的方法准确、安全,应用于有先证者遗传信息的SMA NIPT有一定可行性。Objective：To explore the feasibility of noninvasive prenatal testing（NIPT） applying to spinal muscular atrophy（SMA）. Methods：Twenty-four families with SMA proband children and pregnant again were recruited. The parents and proband children were tested for multiple mutations（ MLPA）. Collect peripheral blood of all family members and plasma of women after 11 weeks of pregnancy. Use the target sequence capture and high-throughput sequencing technique to acquire the target region capture sequences of peripheral blood samples from all family members and obtain the parental haplotype information linked to the pathogenic site, and then com- bine the figure of single nucleotide polymorphism provided by plasma sequencing data. Construct a hidden markov model to analyze fetal haplotype and get the fetal genotype. Use simultaneous invasive prenatal diagnosis of ML- PA to verify its accuracy. This study was aimed at the detection of exon 7/8 of SMN1 gene. Results：Through NIPT,5 were diagnosed as in suffer,7 as a carrier and 12 were normal. The results of MLPA test showed that 5 in suffer,8 in carrier and 11 in normal. All the suffering fetuses were homozygous for exon 7 and 8 of SMN1 gene. The coincidence rate of NIPT and MLPA test was 95.83% （23/24）, and the difference had no statistical signifi- cance （ P = 0. 317）. Conclusions： The targeted region capture sequencing and haplotype-based noninvasive prenatal testing for SMA is accurate and may have potential use in clinical applications.

关 键 词：目标区域捕获测序 单体型 脊髓性肌肉萎缩症 无创产前检测 胎儿游离DNA 

分 类 号：R714.5[医药卫生—妇产科学]