Noninvasive Prenatal Diagnosis of Monogenic Diseases by Targeted Massively Parallel Sequencing of Maternal Plasma: Application to β-Thalassemia
Noninvasive Prenatal Diagnosis of Monogenic Diseases by Targeted Massively Parallel Sequencing of Maternal Plasma: Application to β-Thalassemia
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DOI：
10.1373/clinchem.2012.189589
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发表时间：
2012-10-01
影响因子：
9.3
通讯作者：
Lo, Y. M. Dennis
中科院分区：
文献类型：
--
作者：
Lam, Kwan-Wood G.;Jiang, Peiyong;Lo, Y. M. Dennis
背景：最近通过对母体血浆DNA的深度测序，确定了一个胎儿的全基因组遗传和突变图谱。这项技术可能在许多单基因疾病的无创性产前诊断(NIPD)中有重要应用。相对单倍型剂量(RHDO)分析是这一过程的核心步骤，它将使人们能够阐明胎儿基因组中母亲遗传的一半。对于临床应用，通过有针对性地将这种方法应用于包含致病基因的选定基因组区域，可以降低数据分析的成本和复杂性。因此，有必要探索以有针对性的方式进行RHDO分析的可行性。方法：我们在2个接受β-地中海贫血产前诊断的家庭中，通过溶液相杂交和大规模平行测序进行了靶向浓缩。我们使用数字聚合酶链式反应策略来物理推断父母的单倍型。结果：通过对母体血浆DNA进行靶向测序，获得了β-珠蛋白基因区域206倍的平均测序深度。RHDO分析成功地分析了从靶浓缩样本获得的测序数据，包括其中一个家庭中父母具有相似单倍型结构的区域。数据分析表明，两个胎儿都是β-地中海贫血的杂合子携带者。结论：对单基因疾病的NIPD进行母体血浆DNA靶向测序是可行的。(C)2012年美国临床化学协会
BACKGROUND: A genomewide genetic and mutational profile of a fetus was recently determined via deep sequencing of maternal plasma DNA. This technology could have important applications for noninvasive prenatal diagnosis (NIPD) of many monogenic diseases. Relative haplotype dosage (RHDO) analysis, a core step of this procedure, would allow one to elucidate the maternally inherited half of the fetal genome. For clinical applications, the cost and complexity of data analysis might be reduced via targeted application of this approach to selected genomic regions containing disease-causing genes. There is thus a need to explore the feasibility of performing RHDO analysis in a targeted manner.METHODS: We performed target enrichment by using solution-phase hybridization followed by massively parallel sequencing of the beta-globin gene region in 2 families undergoing prenatal diagnosis for beta-thalassemia. We used digital PCR strategies to physically deduce parental haplotypes. Finally, we performed RHDO analysis with target-enriched sequencing data and parental haplotypes to reveal the beta-thalassemic status for the fetuses.RESULTS: A mean sequencing depth of 206-fold was achieved in the beta-globin gene region by targeted sequencing of maternal plasma DNA. RHDO analysis was successful for the sequencing data obtained from the target-enriched samples, including a region in one of the families in which the parents had similar haplotype structures. Data analysis revealed that both fetuses were heterozygous carriers of beta-thalassemia.CONCLUSIONS: Targeted sequencing of maternal plasma DNA for NIPD of monogenic diseases is feasible. (C) 2012 American Association for Clinical Chemistry