A New Era for PGD Testing: One Novel All-Encompassing Diagnostic Method
AWARD NOMINEE

Introduction:
Recent technological advances have enabled combined gene mutation detection and comprehensive chromosome screening (CCS) from a single embryo biopsy by means of microarray or high coverage next generation sequencing (NGS) technologies. However, these methods are still challenging, very expensive, time consuming, and therefore not widely applied.

Aim:
To develop an all-in-one NGS-based workflow for gene mutation detection combined with CCS screening at a third of the cost of competing array and high throughput sequencing-based technologies while maintaining the highest standard for accuracy in a 24 hour workflow protocol.

Materials & Methods:
Single lymphoblast cells from tissue cultures of a family of 4 siblings with BRCA2 gene mutation carriage were subjected to low coverage (1x) whole genome sequencing on a NextSeq 500 instrument followed by combined chromosome copy number assessment and whole genome haplotype phasing. Validation of both haplotype and copy number predictions in single cell data was performed by SNP microarray analysis of bulk DNA from each of the tested siblings.

Results:
Copy number assessment and haplotype phasing of the single cell sequencing data in the BRCA2 gene region and the entire genome was completely (100%) concordant with both known BRCA2 genotypes and microarray determined whole genome chromosome copy numbers and haplotypes in all siblings.

Conclusions:
Our results demonstrate the establishment of a reliable method for all-in-one molecular and chromosomal diagnosis of single cells. Our novel NGS –based method may become the ultimate, rapid, low cost solution for combined PGD and CCS testing.