title: Methods to detect CNVs in the human genome.
abstract: The detection of quantitative changes in genomic DNA, i.e. deletions and duplications or Copy Number Variants (CNVs), has recently gained considerable interest. First, detailed analysis of the human genome showed a surprising amount of CNVs, involving thousands of genes. Second, it was realised that the detection of CNVs as a cause of genetic disease was often neglected, but should be an essential part of a complete screening strategy. In both cases new efficient CNV screening methods, covering the entire range from specific loci to genome-wide, were behind these developments. This paper will briefly review the methods that are available to detect CNVs, discuss their strong and weak points, show some new developments and look ahead. Methods covered include microscopy, fluorescence in situ hybridization (including fiber-FISH), Southern blotting, PCR-based methods (including MLPA), array technology and massive parallel sequencing. In addition, we will show some new developments, including a 1400-plex CNV bead assay, fast-MLPA (from DNA to result in approximately 6 h) and a simple Melting Curve Analysis assay to confirm potential CNVs. Using the 1400-plex CNV bead assay, targeting selected chromosomal regions only, we detected confirmed rearrangements in 9% of 320 mental retardation patients studied.
authors.affiliation: Human and Clinical Genetics and Leiden Genome Technology Center (LGTC), Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: E
authors.initials: E
authors.lastname: Aten
authors.forename: S J
authors.initials: SJ
authors.lastname: White
authors.forename: M E
authors.initials: ME
authors.lastname: Kalf
authors.forename: R H A M
authors.initials: RH
authors.lastname: Vossen
authors.forename: H H
authors.initials: HH
authors.lastname: Thygesen
authors.forename: C A
authors.initials: CA
authors.lastname: Ruivenkamp
authors.forename: M
authors.initials: M
authors.lastname: Kriek
authors.forename: M H B
authors.initials: MH
authors.lastname: Breuning
authors.forename: J T
authors.initials: JT
authors.lastname: den Dunnen
mesh_terms: D018628:Gene Dosage / Q000235:genetics*; D005821:Genetic Techniques; D015894:Genome, Human / Q000235:genetics*; D006801:Humans; D013997:Time Factors