Reliable detection of subchromosomal deletions and duplications using cell-based noninvasive prenatal testing

Vossaert, LiesbethWang, QunSalman, RoseenZhuo, XinmingQu, ChunjingHenke, DavidSeubert, RonChow, JenniferU'ren, LanceEnright, BrennanStilwell, JackieKaldjian, EricYang, YapingShaw, ChadLevy, BrynnWapner, RonaldBreman, AmyVeyver, Ignatia Van denBeaudet, Arthur2019-06-142019-06-142018-10-25Vossaert, L., Wang, Q., Salman, R., Zhuo, X., Qu, C., Henke, D., … Beaudet, A. (2018). Reliable detection of subchromosomal deletions and duplications using cell-based noninvasive prenatal testing. Prenatal Diagnosis, 38(13), 1069–1078. https://doi.org/10.1002/pd.53771097-0223https://hdl.handle.net/1805/19618Objective To gather additional data on the ability to detect subchromosomal abnormalities of various sizes in single fetal cells isolated from maternal blood, using low-coverage shotgun next-generation sequencing for cell-based noninvasive prenatal testing (NIPT). Method Fetal trophoblasts were recovered from approximately 30 mL of maternal blood using maternal white blood cell depletion, density-based cell separation, immunofluorescence staining, and high-resolution scanning. These trophoblastic cells were picked as single cells and underwent whole genome amplification for subsequent genome-wide copy number analysis and genotyping to confirm the fetal origin of the cells. Results Applying our fetal cell isolation method to a series of 125 maternal blood samples, we detected on average 4.17 putative fetal cells/sample. The series included 15 cases with clinically diagnosed fetal aneuploidies and five cases with subchromosomal abnormalities. This method was capable of detecting findings that were 1 to 2 Mb in size, and all were concordant with the microarray or karyotype data obtained on a fetal sample. A minority of fetal cells showed evidence of genome degradation likely related to apoptosis. Conclusion We demonstrate that this cell-based NIPT method has the capacity to reliably diagnose fetal chromosomal abnormalities down to 1 to 2 Mb in size.en-USAttribution 3.0 United Statessubchromosomal abnormalitiesprenatal testingtrophoblastsReliable detection of subchromosomal deletions and duplications using cell-based noninvasive prenatal testingArticle