一种具有单碱基分辨率的新型高通量分子计数方法可实现精确的单基因 NIPT。
A novel high-throughput molecular counting method with single base-pair resolution enables accurate single-gene NIPT.
机构信息
BillionToOne Inc., Menlo Park, CA, 94025, USA.
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA, 94158, USA.
出版信息
Sci Rep. 2019 Oct 7;9(1):14382. doi: 10.1038/s41598-019-50378-8.
Next-generation DNA sequencing is currently limited by an inability to accurately count the number of input DNA molecules. Molecular counting is particularly needed when accurate quantification is required for diagnostic purposes, such as in single gene non-invasive prenatal testing (sgNIPT) and liquid biopsy. We developed Quantitative Counting Template (QCT) molecular counting to reconstruct the number of input DNA molecules using sequencing data. We then used QCT molecular counting to develop sgNIPTs of sickle cell disease, cystic fibrosis, spinal muscular atrophy, alpha-thalassemia, and beta-thalassemia. The analytical sensitivity and specificity of sgNIPT was >98% and >99%, respectively. Validation of sgNIPTs was further performed with maternal blood samples collected during pregnancy, and sgNIPTs were 100% concordant with newborn follow-up.
下一代 DNA 测序目前受到无法准确计数输入 DNA 分子数量的限制。当需要进行准确的定量分析以用于诊断目的时，例如在单基因无创产前检测 (sgNIPT) 和液体活检中，分子计数就显得尤为重要。我们开发了定量计数模板 (QCT) 分子计数法，利用测序数据来重建输入 DNA 分子的数量。然后，我们使用 QCT 分子计数法开发了镰状细胞病、囊性纤维化、脊髓性肌萎缩症、α-地中海贫血和β-地中海贫血的 sgNIPT。sgNIPT 的分析灵敏度和特异性分别大于 98%和大于 99%。进一步使用怀孕期间采集的母亲血液样本对 sgNIPT 进行验证，sgNIPT 与新生儿随访结果完全一致。
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