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

发布时间：2018-04-14 18:56

本文选题：目标区域捕获测序 + 单体型　；参考：《实用妇产科杂志》2017年09期

【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。
[Abstract]:Objective: to explore the feasibility of non-invasive prenatal examination (NIPT) in spinal muscular dystrophy (Spinal muscular dystrophy).Methods: a total of 24 families with SMA probands were recruited for further pregnancy. The pathogenic mutations of both husband and wife and proband were detected by multiplex probe amplification technique.The peripheral blood and maternal plasma after 11 weeks of gestation were collected. The target region of DNA samples from both husband and wife and proband were sequenced by target sequence capture and high-throughput sequencing technique.The haplotype information linked to the pathogenic loci was obtained. Combining with the analysis results of the single nucleotide polymorphism sites provided by plasma sequencing data, a hidden Markov model was constructed to infer the fetal haplotypes, and the fetal genotypes were obtained.At the same time, the accuracy of MLPA was verified by invasive prenatal diagnosis.This study focused on the detection of exons 7 and 8 of SMN1 gene.Results: 5 cases of infected fetus and 7 cases of carrier were detected by NIPT in 24 families.The rate of homozygous deletion of exon 7 and exon 8 of SMN1 gene. NIPT was consistent with that of MLPA was 95.833 / 24. There was no significant difference in diagnostic results between the two methods.Conclusion: based on maternal plasma target region capture and sequencing, haplotype analysis of fetal SMA risk in NIPT is accurate and safe. It is feasible to apply it to SMA NIPT with proband genetic information.
【作者单位】：广州医科大学附属第三医院广州妇产科研究所;深圳华大基因研究院;天津华大基因研究院翻译组学中心;广州华大基因研究院;
【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)
【分类号】：R714.5

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