The Value of Parental Testing by Next-Generation Sequencing Includes the Detection of Germline Mosaicism
The Value of Parental Testing by Next-Generation Sequencing Includes the Detection of Germline Mosaicism
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DOI：
10.1016/j.jmoldx.2020.02.001
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发表时间：
2020-05-01
影响因子：
4.1
通讯作者：
Strom, Samuel P.
中科院分区：
文献类型：
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作者：
Brewer, Casey J.;Gillespie, Meghan;Strom, Samuel P.
当在先证者中检测到潜在的致病变异时，父母测试用于确定遗传模式。这项研究表明，下一代测序（NGS）是非常适合父母的测试，特别是因为它能够检测临床相关的生殖系嵌合。在临床实验室中通过NGS进行亲本变异检测1年。将通过NGS检测嵌合体与通过桑格测序检测嵌合体进行比较。NGS在8个不同的基因中检测到8例先前未发现的嵌合体。使用定制生物信息学分析结合家族遗传数据和互补桑格测序将镶嵌变体与测序噪声区分开。桑格测序通过NGS检测到等位基因分数>= 8%的嵌合变体，但不能检测到低于该水平的嵌合变体。通过NGS检测生殖系嵌合体对父母来说是非常宝贵的，提供了更准确的复发风险，可以改变计划生育和妊娠管理的决定。由于NGS还可以确认亲子关系并提高可扩展性，因此它同时简化并加强了变体策展过程。这些特征使得NGS成为亲本检测的理想方法，对于大多数基因组位点，甚至比桑格测序上级。
When a potential disease-causing variant is detected in a proband, parental testing is used to determine the mode of inheritance. This study demonstrates that next-generation sequencing (NGS) is uniquely well suited for parental testing, in particular because of its ability to detect clinically relevant germline mosaicism. Parental variant testing by NGS was performed in a clinical laboratory for 1 year. The detection of mosaicism by NGS was compared with its detection by Sanger sequencing. Eight cases of previously unrevealed mosaicism were detected by NGS across eight different genes. Mosaic variants were differentiated from sequencing noise using custom bioinformatics analyses in combination with familial inheritance data and complementary Sanger sequencing. Sanger sequencing detected mosaic variants with allele fractions >= 8% by NGS, but could not detect mosaic variants below that level. Detection of germline mosaicism by NGS is invaluable to parents, providing a more accurate recurrence risk that can alter decisions on family planning and pregnancy management. Because NGS can also confirm parentage and increase scalability, it simultaneously streamlines and strengthens the variant curation process. These features make NGS the ideal method for parental testing, superior even to Sanger sequencing for most genomic loci.