RT Journal Article
SR Electronic
T1 A novel high-throughput molecular counting method with single base-pair resolution enables accurate single-gene NIPT
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 597732
DO 10.1101/597732
A1 David S. Tsao
A1 Sukrit Silas
A1 Brian P. Landry
A1 Nelda Itzep
A1 Amy B. Nguyen
A1 Celeste K. Kanne
A1 Vivien A. Sheehan
A1 Rani Sharma
A1 Rahul Shukla
A1 Prem N. Arora
A1 Oguzhan Atay
YR 2019
UL http://biorxiv.org/content/early/2019/04/03/597732.abstract
AB Next-generation DNA sequencing is currently limited by an inability to 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 for reconstructing the number of input DNA molecules using sequencing data. We then used QCT molecular counting to develop sgNIPT of sickle cell disease, cystic fibrosis, spinal muscular atrophy, alpha-thalassemia, and beta-thalassemia. Incorporating molecular count information into a statistical model of disease likelihood led to analytical sensitivity and specificity of >98% and >99%, respectively. Validation of sgNIPT was further performed with maternal blood samples collected during pregnancy, and sgNIPT was 100% concordant with newborn follow-up.