Validation study for noninvasive single-cell-based prenatal genetic testing

Objective To clinically validate a cell-based noninvasive prenatal genetic test using sequence-based copy number analysis of single trophoblasts from maternal blood. Methods Blood was obtained from 401 individuals (8-22 weeks) and shipped overnight. Red blood cells were lysed, and nucleated cells stained for cytokeratin (CK) and CD45 using fluorescent antibodies and enriched for positive CK staining. Automated microscopic scanning was used to identify and pick single CK+/CD45- trophoblasts which were subjected to whole genome amplification and next-generation sequencing. Results Blood was obtained from 243 pregnancies scheduled for CVS or amniocentesis. Luna results were normal for 160 singletons while 15 cases were abnormal (14 aneuploidy and one monozygotic twin case with Williams syndrome deletion). These Luna results agreed with CVS/amniocentesis. Placental mosaicism occurred in 7 of 236 (3.0%) Luna cases and in 3 of 188 (1.6%) CVS cases (total 4.6%). No scorable trophoblasts were recovered in 32 of 236 (13.6%) usable samples. Additionally, 158 low-risk pregnancies not undergoing CVS/amniocentesis showed normal results for 133 cases. Seven had aneuploidy results, and there were 3 likely pathogenic deletions or duplications including one15q11-q13 deletion. Conclusion This noninvasive cell-based prenatal genetic test detected aneuploidy and deletions/duplications with high sensitivity and specificity based on concordance with CVS/amniocentesis. What’s already known about this topic? What does this study add? ### Competing Interest Statement All authors based at Luna Genetics are or were Luna Genetics employees. ALB is a founder and significant stockholder for Luna Genetics. No other conflicts of interest. ### Funding Statement This study was funded by Luna Genetics, Inc. ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: Columbia Medical Center - Columbia University IRB 5. Protocol Number: IRB-AAAQ8062 Level of review and outcome: Approved by Expedited review. December 9, 2019 PI:Ronald Wapner 7528304 - OBG Rsch MFM Rsch Protocol Number: IRB-AAAQ8062 WCG IRB (Formerly Western IRB) PI Beaudet IRB Tracking Number: 20193442 Board Action Date: 01/04/2023 I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes