Custom Array Comparative Genomic Hybridization: the Importance of DNA Quality, an Expert Eye, and Variant Validation

• Published in: International journal of molecular sciences Vol. 18; no. 3; p. 609
• Main Authors: Lantieri, Francesca, Malacarne, Michela, Gimelli, Stefania, Santamaria, Giuseppe, Coviello, Domenico, Ceccherini, Isabella
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI 10.03.2017
• Subjects: Comparative Genomic Hybridization - methods; Comparative Genomic Hybridization - standards; DNA - standards; Humans; Observer Variation; Signal-To-Noise Ratio; Software; CNV detection filters; agilent aberration call software; high density custom CGH array
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms18030609

The presence of false positive and false negative results in the Array Comparative Genomic Hybridization (aCGH) design is poorly addressed in literature reports. We took advantage of a custom aCGH recently carried out to analyze its design performance, the use of several Agilent aberrations detection algorithms, and the presence of false results. Our study provides a confirmation that the high density design does not generate more noise than standard designs and, might reach a good resolution. We noticed a not negligible presence of false negative and false positive results in the imbalances call performed by the Agilent software. The Aberration Detection Method 2 (ADM-2) algorithm with a threshold of 6 performed quite well, and the array design proved to be reliable, provided that some additional filters are applied, such as considering only intervals with average absolute log2ratio above 0.3. We also propose an additional filter that takes into account the proportion of probes with log2ratio exceeding suggestive values for gain or loss. In addition, the quality of samples was confirmed to be a crucial parameter. Finally, this work raises the importance of evaluating the samples profiles by eye and the necessity of validating the imbalances detected.