Diagnosis of copy number variation by Illumina next generation sequencing is comparable in performance to oligonucleotide array comparative genomic hybridisation

Genomics. 2013 Sep;102(3):174-81. doi: 10.1016/j.ygeno.2013.04.006. Epub 2013 Apr 15.


Array comparative genomic hybridisation (aCGH) profiling is currently the gold standard for genetic diagnosis of copy number. Next generation sequencing technologies provide an alternative and adaptable method of detecting copy number by comparing the number of sequence reads in non-overlapping windows between patient and control samples. Detection of copy number using the BlueGnome 8×60k oligonucleotide aCGH platform was compared with low resolution next generation sequencing using the Illumina GAIIx on 39 patients with developmental delay and/or learning difficulties who were referred to the Leeds Clinical Cytogenetics Laboratory. Sensitivity and workflow of the two platforms were compared. Customised copy number algorithms assessed sequence counts and detected changes in copy number. Imbalances detected on both platforms were compared. Of the thirty-nine patients analysed, all eleven imbalances detected by array CGH and confirmed by FISH or Q-PCR were also detected by CNV-seq. In addition, CNV-seq reported one purported pathogenic copy number variant that was not detected by array CGH. Non-pathogenic, unconfirmed copy number calls were detected by both platforms; however few were concordant between the two. CNV-seq offers an alternative to array CGH for copy number analysis with resolution and future costs comparable to conventional array CGH platforms and with less stringent sample requirements.

Keywords: CNV; Copy number; Expectation–maximization; E–M; FISH; GaII x; Illumina; Illumina Genome Analyzer IIx; MLPA; NGS; Next generation sequencing; OMIM; Online Mendelian Inheritance in Man; Q-PCR; Quantitative PCR; copy number variation; fluorescent in situ hybridisation; multiplex ligation-dependent probe amplification; next generation sequencing.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Child
  • Child, Preschool
  • Chromosome Aberrations
  • Comparative Genomic Hybridization*
  • DNA Copy Number Variations*
  • Genome, Human
  • High-Throughput Nucleotide Sequencing*
  • Humans
  • In Situ Hybridization, Fluorescence
  • Oligonucleotide Array Sequence Analysis*
  • Statistics as Topic