title: Assessment of the frequency of allelic imbalance in human tissue using a multiplex polymerase chain reaction system.
abstract: Genomic instability can generate chromosome breakage and fusion randomly throughout the genome, frequently resulting in allelic imbalance, a deviation from the normal 1:1 ratio of maternal and paternal alleles. Allelic imbalance reflects the karyotypic complexity of the cancer genome. Therefore, it is reasonable to speculate that tissues with more sites of allelic imbalance have a greater likelihood of having disruption of any of the numerous critical genes that cause a cancerous phenotype and thus may have diagnostic or prognostic significance. For this reason, it is desirable to develop a robust method to assess the frequency of allelic imbalance in any tissue. To address this need, we designed an economical and high-throughput method, based on the Applied Biosystems AmpFlSTR Identifiler multiplex polymerase chain reaction system, to evaluate allelic imbalance at 16 unlinked, microsatellite loci located throughout the genome. This method provides a quantitative comparison of the extent of allelic imbalance between samples that can be applied to a variety of frozen and archival tissues. The method does not require matched normal tissue, requires little DNA (the equivalent of approximately 150 cells) and uses commercially available reagents, instrumentation, and analysis software. Greater than 99% of tissue specimens with >or=2 unbalanced loci were cancerous.
authors: Department of Biochemistry and Molecular Biology, MSC08 4670, 1 University of New Mexico, Albuquerque, NM 87131-0001, USA.
authors: Christopher M
authors: CM
authors: Heaphy
authors: William C
authors: WC
authors: Hines
authors: Kimberly S
authors: KS
authors: Butler
authors: Christina M
authors: CM
authors: Haaland
authors: Glenroy
authors: G
authors: Heywood
authors: Edgar G
authors: EG
authors: Fischer
authors: Marco
authors: M
authors: Bisoffi
authors: Jeffrey K
authors: JK
authors: Griffith
mesh_terms: D000483:Alleles; D022981:Allelic Imbalance / Q000235:genetics*; D002292:Carcinoma, Renal Cell / Q000235:genetics; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics; D009369:Neoplasms / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results
chemical_list: D004273:DNA, Neoplasm