Oligonucleotide arrays vs. metaphase-comparative genomic hybridisation and BAC arrays for single-cell analysis: first applications to preimplantation genetic diagnosis for Robertsonian translocation carriers

• Published in: PloS one 2014-11, Vol.9 (11), p.e113223-e113223
• Main Authors: Ramos, Laia, del Rey, Javier, Daina, Gemma, García-Aragonés, Manel, Armengol, Lluís, Fernandez-Encinas, Alba, Parriego, Mònica, Boada, Montserrat, Martinez-Passarell, Olga, Martorell, Maria Rosa, Casagran, Oriol, Benet, Jordi, Navarro, Joaquima
• Format: Article
• Language: English
• Subjects: Artificial chromosomes; Bacterial artificial chromosomes; Biology and Life Sciences; Blastomeres; Blastomeres - cytology; Blastomeres - metabolism; Carriers; Cell Line; Chromosome Banding; Chromosome Disorders - diagnosis; Chromosome Disorders - genetics; Chromosomes, Artificial, Bacterial - genetics; Cleavage; Comparative analysis; Comparative Genomic Hybridization - methods; Cytogenetics; Deoxyribonucleic acid; Diagnosis; DNA; DNA microarrays; Embryos; Female; Fibroblasts; Genetic screening; Genomes; Humans; Hybridization; Karyotyping; Laboratories; Medical diagnosis; Medicine and Health Sciences; Metaphase; Metaphase - genetics; Oligonucleotide Array Sequence Analysis - methods; Oligonucleotides; Pregnancy; Preimplantation Diagnosis - methods; Reproducibility of Results; Robertsonian translocation; Sensitivity and Specificity; Single-Cell Analysis - methods; Sperm; Translocation; Translocation, Genetic
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0113223

Comprehensive chromosome analysis techniques such as metaphase-Comparative Genomic Hybridisation (CGH) and array-CGH are available for single-cell analysis. However, while metaphase-CGH and BAC array-CGH have been widely used for Preimplantation Genetic Diagnosis, oligonucleotide array-CGH has not been used in an extensive way. A comparison between oligonucleotide array-CGH and metaphase-CGH has been performed analysing 15 single fibroblasts from aneuploid cell-lines and 18 single blastomeres from human cleavage-stage embryos. Afterwards, oligonucleotide array-CGH and BAC array-CGH were also compared analysing 16 single blastomeres from human cleavage-stage embryos. All three comprehensive analysis techniques provided broadly similar cytogenetic profiles; however, non-identical profiles appeared when extensive aneuploidies were present in a cell. Both array techniques provided an optimised analysis procedure and a higher resolution than metaphase-CGH. Moreover, oligonucleotide array-CGH was able to define extra segmental imbalances in 14.7% of the blastomeres and it better determined the specific unbalanced chromosome regions due to a higher resolution of the technique (≈ 20 kb). Applicability of oligonucleotide array-CGH for Preimplantation Genetic Diagnosis has been demonstrated in two cases of Robertsonian translocation carriers 45,XY,der(13;14)(q10;q10). Transfer of euploid embryos was performed in both cases and pregnancy was achieved by one of the couples. This is the first time that an oligonucleotide array-CGH approach has been successfully applied to Preimplantation Genetic Diagnosis for balanced chromosome rearrangement carriers.