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(2025-12-31 22:50):
#paper doi: https://doi.org/10.1038/s41598-019-50378-8. Scientific Reports. 2019. A novel high-throughput molecular counting method with single base-pair resolution enables accurate single-gene NIPT. 在NIPT技术应用中，无论是传统的无创检测还是近年来不断发展的无创单基因病检测，分子计数非常关键。这篇文章开发一个叫做 Quantitative Counting Template (QCT)的分子计数技术。简单说就是测在扩增和测序序之前，在模板DNA上添加了一个Embedded Molecular Index (EMI)独特分子标签，然后再进行扩增和测序，在获得测序数据后，通过EMI来识别模板分子，进而实现更为准确的计数。随后，研究人员基于这个技术，开发了针对镰状细胞病、囊性纤维化、脊髓性肌萎缩症、α地中海贫血及β地中海贫血（sickle cell disease, cystic fibrosis, spinal muscular atrophy, alpha-thalassemia, and beta-thalassemia）的单基因NIPT（sgNIPT）检测。该检测的分析敏感性与特异性均超过98%和99%。通过妊娠期采集的母体血液样本进一步验证了sgNIPT检测，其结果与新生儿随访检测100%一致。近年来，无创单基因病检测技术日渐增多和成熟，但直观感受上来看，单基因病的NIPT检测更多地要依靠实验环节的技术创新来达成。
Scientific Reports,
2019-10-7.
DOI: 10.1038/s41598-019-50378-8
A novel high-throughput molecular counting method with single base-pair resolution enables accurate single-gene NIPT
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Abstract:
Abstract 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.
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