Use of amplicon-based sequencing for testing fetal identity and monogenic traits with Single Circulating Trophoblast (SCT) as one form of cell-based NIPT

A major challenge for cell-based non-invasive prenatal testing (NIPT) is to distinguish individual presumptive fetal cells from maternal in female pregnancies. We have sought a rapid, robust, versatile, and low-cost next-generation sequencing method facilitate this process. Toward goal, single isolated underwent whole genome amplification prior genotyping. Multiple highly polymorphic genomic regions (including HLA-A HLA-B) with 10–20 very informative nucleotide polymorphisms (SNPs) within 200 bp interval were amplified modified based on other publications. To enhance the power of cell identification, approximately 40 Human Identification SNP (Applied Biosystems) test amplicons also utilized. Using results compare sex chromosome data NGS as reliable standard, true positive rate genotyping was 83.4%, negative 6.6%, false 3.3%, 6.6%. These would not be sufficient clinical diagnosis, but they demonstrate general validity approach suggest that deeper could completely reliable. paternal DNA sample required using method. The assay successfully detected pathogenic variants causing Tay Sachs disease, cystic fibrosis, hemoglobinopathies lymphoblastoid cells, disease-causing three NIPT cases. This applicable any monogenic diagnosis.