Non-invasive prenatal diagnosis of spinal muscular atrophy by relative haplotype dosage.
Non-invasive prenatal diagnosis of spinal muscular atrophy by relative haplotype dosage.
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DOI：
10.1038/ejhg.2016.195
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发表时间：
2017-04
期刊：
影响因子：
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通讯作者：
Allen S
中科院分区：
文献类型：
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作者：
Parks M;Court S;Bowns B;Cleary S;Clokie S;Hewitt J;Williams D;Cole T;MacDonald F;Griffiths M;Allen S
虽然技术上是可行的，但世界各地很少有临床实验室对选定的单基因疾病实施非侵入性产前诊断（NIPD），主要是由于成本上升。先前已经证明X连锁疾病的NIPD可以在临床实践中可行地实施，我们现在已经开发了一种常染色体隐性遗传疾病脊髓性肌萎缩症（SMA）的NIPD测试。从母体血液中提取无细胞DNA，并制备用于在Illumina MiSeq上进行大规模平行测序，通过在含有SMN1基因的5号染色体上的6 Mb基因组窗口上靶向捕获富集单核苷酸多态性。母亲，父亲和先证者的DNA样本也进行了测试的单倍型的目的。测序数据通过相对单倍型剂量（RHDO）进行分析。通过NIPSIGEN研究招募了6名妊娠SMA携带者和10名健康妊娠供体。通过RHDO常染色体隐性遗传疾病分析，确定胎儿中受影响SMN1基因的母系和父系衍生等位基因的遗传。先证者（SMA携带者）或侵入性获得的胎儿样本（健康妊娠供体）的DNA用于鉴定与受影响SMN1基因相关的母体和父亲参考单倍型。所有患者的结果与已知结果相关，并显示检测的特异性和灵敏度为100%。除了在整个验证阶段显示出高准确性和可靠性之外，我们的SMA NIPD新测试也是经济实惠的，并且可以应用于临床服务。
Although technically possible, few clinical laboratories across the world have implemented non-invasive prenatal diagnosis (NIPD) for selected single-gene disorders, mostly owing to the elevated costs incurred. Having previously proven that NIPD for X-linked disorders can be feasibly implemented in clinical practice, we have now developed a test for the NIPD of an autosomal-recessive disorder, spinal muscular atrophy (SMA). Cell-free DNA was extracted from maternal blood and prepared for massively parallel sequencing on an Illumina MiSeq by targeted capture enrichment of single-nucleotide polymorphisms across a 6 Mb genomic window on chromosome 5 containing the SMN1 gene. Maternal, paternal and proband DNA samples were also tested for haplotyping purposes. Sequencing data was analysed by relative haplotype dosage (RHDO). Six pregnant SMA carriers and 10 healthy pregnant donors were recruited through the NIPSIGEN study. Inheritance of the maternally and paternally derived alleles of the affected SMN1 gene was determined in the foetus by RHDO analysis for autosomal-recessive disorders. DNA from the proband (for SMA carriers) or an invasively obtained foetal sample (for healthy pregnant donors) was used to identify the maternal and paternal reference haplotypes associated with the affected SMN1 gene. Results for all patients correlated with known outcomes and showed a testing specificity and sensitivity of 100%. On top of showing high accuracy and reliability throughout the stages of validation, our novel test for NIPD of SMA is also affordable and viable for implementation into clinical service.