Background Non-invasive prenatal testing (NIPT) has been widely adopted for the detection of fetal aneuploidies and microdeletion syndromes, nevertheless, limited clinical utilization has been reported for the non-invasive prenatal screening of monogenic diseases. In this study, we present the development and validation of a single comprehensive NIPT for prenatal screening of chromosomal aneuploidies, microdeletions and 50 autosomal recessive disorders associated with severe or moderate clinical phenotype. Results We employed a targeted capture enrichment technology powered by custom TArget Capture Sequences (TACS) and multi-engine bioinformatics analysis pipeline to develop and validate a novel NIPT test. This test was validated using 2033 cell-fee DNA (cfDNA) samples from maternal plasma of pregnant women referred for NIPT and paternal genomic DNA. Additionally, 200 amniotic fluid and CVS samples were used for validation purposes. All NIPT samples were correctly classified exhibiting 100% sensitivity (CI 89.7-100%) and 100% specificity (CI 99.8-100%) for chromosomal aneuploidies and microdeletions. Furthermore, 613 targeted causative mutations, of which 87 were unique, corresponding to 21 monogenic diseases, were identified. For the validation of the assay for prenatal diagnosis purposes, all aneuploidies, microdeletions and point mutations were correctly detected in all 200 amniotic fluid and CVS samples. Conclusions We present a NIPT for aneuploidies, microdeletions, and monogenic disorders. To our knowledge this is the first time that such a comprehensive NIPT is available for clinical implementation.

中文翻译：

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靶向捕获富集，然后进行 NGS：开发和验证针对染色体非整倍体、微缺失综合征和单基因疾病的单一综合 NIPT。


背景 无创产前检测（NIPT）已广泛用于检测胎儿非整倍体和微缺失综合征，然而，单基因疾病无创产前筛查的临床应用有限。在本研究中，我们开发并验证了单一综合 NIPT，用于产前筛查染色体非整倍体、微缺失和 50 种与重度或中度临床表型相关的常染色体隐性遗传疾病。结果我们采用了由定制目标捕获序列 (TACS) 和多引擎生物信息学分析流程支持的靶向捕获富集技术来开发和验证新型 NIPT 测试。该测试使用 2033 个来自孕妇血浆的无细胞 DNA (cfDNA) 样本和父本基因组 DNA 进行验证。此外，还使用 ​​200 份羊水和 CVS 样本进行验证。所有 NIPT 样本均经过正确分类，对染色体非整倍体和微缺失表现出 100% 敏感性 (CI 89.7-100%) 和 100% 特异性 (CI 99.8-100%)。此外，还鉴定出了 613 个靶向致病突变，其中 87 个是独特的，对应 21 种单基因疾病。为了验证产前诊断的检测方法，所有 200 份羊水和 CVS 样本中的所有非整倍体、微缺失和点突变均得到正确检测。结论 我们提出了针对非整倍体、微缺失和单基因疾病的 NIPT。据我们所知，这是第一次将如此全面的 NIPT 应用于临床。