Quantitative PCR evaluation of deletions/duplications identified by array CGH. (August 2019)

• Record Type: Journal Article
• Authors: Baldan, Federica; Passon, Nadia; Burra, Silvia; Demori, Eliana; Russo, Patrizia Dello; Damante, Giuseppe
• Is Part Of: Molecular and cellular probes. Volume 46(2019)
• Publication Date: 2019-08
• Subjects: Array CGH -- Quantitative PCR -- CNV -- Deletion -- Duplication; Molecular probes -- Diagnostic use -- Periodicals; Pathology, Cellular -- Technique -- Periodicals; Cell Biology -- Periodicals; Molecular Biology -- Periodicals; Sondes moléculaires -- Utilisation diagnostique -- Périodiques; Cytopathologie -- Technique -- Périodiques; 572
• DOI: 10.1016/j.mcp.2019.101421
• Languages: English
• ISSNs: 0890-8508

Abstract

Abstract: Genomic deletions/duplications detected by array comparative genomic hybridization (aCGH) should be confirmed by an independent technology. This approach allows also to test, at low cost, inheritance of the imbalance. In the present study we explored the use of quantitative PCR (qPCR) to confirm aCGH-detected potentially clinically relevant imbalances. Only samples with DLRS <0.2 were tested for confirmation. aCGH results were confirmed in 102/118 cases (86.5%). A major element for non-confirmation was the dimension (and the probe coverage) of the putative aberration. Imbalances detected by 10 or less probes in aCGH assay were not confirmed in 11 out of 41 cases (26.8%), while those ones detected by 20 or more probes were always confirmed (46 cases). Among not confirmed imbalances, no statistical difference was found between deletions and duplication. Our data indicate that validation should be required for imbalances detected by less than 10 probes in aCGH assays. Highlights: Quantitative PCR has been used to evaluate deletions or duplications identified by array CGH. Amplimers were generated to validate presence of aberration for 118 genomic regions and confirmation was obtained in 86.5% of cases. A major element for non-confirmation was the dimension (and the probes coverage) of the putative aberration.