title: Sample degradation leads to false-positive copy number variation calls in multiplex real-time polymerase chain reaction assays.
abstract: The recent implication of genomic copy number variations (CNVs) in multiple human genetic disorders has led to increased interest in CNV discovery technologies. There is a growing consensus that, in addition to the method used for detection, at least one additional technology should be employed for validation. Real-time quantitative polymerase chain reaction (qPCR) analysis, incorporating a normal (2N) copy number standard, is commonly used as a means of validating CNVs. Whereas it has previously been reported that formalin-fixed paraffin-embedded (FFPE) DNA samples can yield spurious CNV calls in real-time qPCR assays, here we report that sample degradation under standard laboratory storage conditions generates a significant increase in false-positive CNV results. Results suggest the possibility of biased degradation among genomic regions and emphasize the need to assess sample integrity immediately prior to real-time qPCR experiments.
authors.affiliation: Miami Institute for Human Genomics, Miller School of Medicine, University of Miami, Miami, FL 33136, USA.
authors.forename: Holly N
authors.initials: HN
authors.lastname: Cukier
authors.forename: Margaret A
authors.initials: MA
authors.lastname: Pericak-Vance
authors.forename: John R
authors.initials: JR
authors.lastname: Gilbert
authors.forename: Dale J
authors.initials: DJ
authors.lastname: Hedges
mesh_terms: D005189:False Positive Reactions; D018628:Gene Dosage; D014644:Genetic Variation; D015894:Genome, Human; D006801:Humans; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity