title: Studying copy number variations using a nanofluidic platform.
abstract: Copy number variations (CNVs) in the human genome are conventionally detected using high-throughput scanning technologies, such as comparative genomic hybridization and high-density single nucleotide polymorphism (SNP) microarrays, or relatively low-throughput techniques, such as quantitative polymerase chain reaction (PCR). All these approaches are limited in resolution and can at best distinguish a twofold (or 50%) difference in copy number. We have developed a new technology to study copy numbers using a platform known as the digital array, a nanofluidic biochip capable of accurately quantitating genes of interest in DNA samples. We have evaluated the digital array's performance using a model system, to show that this technology is exquisitely sensitive, capable of differentiating as little as a 15% difference in gene copy number (or between 6 and 7 copies of a target gene). We have also analyzed commercial DNA samples for their CYP2D6 copy numbers and confirmed that our results were consistent with those obtained independently using conventional techniques. In a screening experiment with breast cancer and normal DNA samples, the ERBB2 gene was found to be amplified in about 35% of breast cancer samples. The use of the digital array enables accurate measurement of gene copy numbers and is of significant value in CNV studies.
authors: Fluidigm Corporation, South San Francisco, CA 94080, USA.
authors: Jian
authors: J
authors: Qin
authors: Robert C
authors: RC
authors: Jones
authors: Ramesh
authors: R
authors: Ramakrishnan
mesh_terms: D001324:Autoantigens / Q000235:genetics; D001943:Breast Neoplasms / Q000235:genetics; D019389:Cytochrome P-450 CYP2D6 / Q000235:genetics; D005260:Female; D018628:Gene Dosage; D018734:Genes, erbB-2; D014644:Genetic Variation; D006801:Humans; D046210:Microfluidic Analytical Techniques; D036103:Nanotechnology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D043262:Ribonuclease P / Q000235:genetics
chemical_list: D001324:Autoantigens; C104646:RPP30 protein, human; D019389:Cytochrome P-450 CYP2D6; D043262:Ribonuclease P