A Cell-free DNA Barcode-Enabled Single-Molecule Test for Noninvasive Prenatal Diagnosis of Monogenic Disorders: Application to β-Thalassemia
A Cell-free DNA Barcode-Enabled Single-Molecule Test for Noninvasive Prenatal Diagnosis of Monogenic Disorders: Application to β-Thalassemia
复制标题
用于单基因疾病无创产前诊断的无细胞 DNA 条形码单分子测试：在 β 地中海贫血中的应用
DOI：
10.1002/advs.201802332
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发表时间：
2019-06-05
期刊：
影响因子：
15.1
通讯作者：
Xu, Xiangmin
中科院分区：
文献类型：
--
作者：
Yang, Xingkun;Zhou, Qinghua;Xu, Xiangmin
在过去的十年里，对常见的非整倍体进行无创性产前检测已经成为常规，但单基因疾病的检测在临床实施中仍然是一个挑战。最近的大多数研究都有内在的局限性，如复杂的程序，缺乏通用性，以及需要事先了解父母的基因类型或单倍型。为了克服这些局限性，开发了一种强大而通用的基于下一代测序的无细胞DNA(CfDNA)等位基因计数系统，称为cfDNA条形码启用的单分子测试(CfBEST)，用于单基因疾病的无创性产前诊断(NIPD)。研究发现，cfBEST在检测cfDNA低丰度突变方面的准确性与液滴数字聚合酶链式反应(DdPCR)相当。CfBEST的分析有效性通过β-地中海贫血检测得到证明，在对143例高危妊娠进行盲法验证研究中，等位基因检测的敏感性为99.19%，特异性为99.92%。由于验证的cfBEST方法可以精确和定量地检测cfDNA中的母胎基因组合，因此它具有用于人类单基因疾病的NIPD的潜力。
Noninvasive prenatal testing of common aneuploidies has become routine over the past decade, but testing of monogenic disorders remains a challenge in clinical implementation. Most recent studies have inherent limitations, such as complicated procedures, a lack of versatility, and the need for prior knowledge of parental genotypes or haplotypes. To overcome these limitations, a robust and versatile next-generation sequencing-based cell-free DNA (cfDNA) allelic molecule counting system termed cfDNA barcode-enabled single-molecule test (cfBEST) is developed for the noninvasive prenatal diagnosis (NIPD) of monogenic disorders. The accuracy of cfBEST is found to be comparable to that of droplet digital polymerase chain reaction (ddPCR) in detecting low-abundance mutations in cfDNA. The analytical validity of cfBEST is evidenced by a beta-thalassemia assay, in which a blind validation study of 143 at-risk pregnancies reveals a sensitivity of 99.19% and a specificity of 99.92% on allele detection. Because the validated cfBEST method can be used to detect maternal-fetal genotype combinations in cfDNA precisely and quantitatively, it holds the potential for the NIPD of human monogenic disorders.