title: Analysis of copy number variation using quantitative interspecies competitive PCR.
abstract: Over recent years small submicroscopic DNA copy-number variants (CNVs) have been highlighted as an important source of variation in the human genome, human phenotypic diversity and disease susceptibility. Consequently, there is a pressing need for the development of methods that allow the efficient, accurate and cheap measurement of genomic copy number polymorphisms in clinical cohorts. We have developed a simple competitive PCR based method to determine DNA copy number which uses the entire genome of a single chimpanzee as a competitor thus eliminating the requirement for competitive sequences to be synthesized for each assay. This results in the requirement for only a single reference sample for all assays and dramatically increases the potential for large numbers of loci to be analysed in multiplex. In this study we establish proof of concept by accurately detecting previously characterized mutations at the PARK2 locus and then demonstrating the potential of quantitative interspecies competitive PCR (qicPCR) to accurately genotype CNVs in association studies by analysing chromosome 22q11 deletions in a sample of previously characterized patients and normal controls.
authors.affiliation: Department of Psychological Medicine, Wales School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, UK. williamsnm@cf.ac.uk
authors.forename: Nigel M
authors.initials: NM
authors.lastname: Williams
authors.forename: Hywel
authors.initials: H
authors.lastname: Williams
authors.forename: Elisa
authors.initials: E
authors.lastname: Majounie
authors.forename: Nadine
authors.initials: N
authors.lastname: Norton
authors.forename: Beate
authors.initials: B
authors.lastname: Glaser
authors.forename: Huw R
authors.initials: HR
authors.lastname: Morris
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Owen
authors.forename: Michael C
authors.initials: MC
authors.lastname: O'Donovan
mesh_terms: D000818:Animals; D002872:Chromosome Deletion; D002892:Chromosomes, Human, Pair 22; D018628:Gene Dosage; D014644:Genetic Variation; D006801:Humans; D009154:Mutation; D002679:Pan troglodytes; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D011110:Polymorphism, Genetic; D012015:Reference Standards; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA / Q000379:methods*; D044767:Ubiquitin-Protein Ligases / Q000235:genetics
chemical_list: D044767:Ubiquitin-Protein Ligases; C111567:parkin protein