title: Blinded study determination of high sensitivity and specificity microchip electrophoresis-SSCP/HA to detect mutations in the p53 gene.
abstract: Knowledge of the genetic changes that lead to disease has grown and continues to grow at a rapid pace. However, there is a need for clinical devices that can be used routinely to translate this knowledge into the treatment of patients. Use in a clinical setting requires high sensitivity and specificity (>97%) in order to prevent misdiagnoses. Single-strand conformational polymorphism (SSCP) and heteroduplex analysis (HA) are two DNA-based, complementary methods for mutation detection that are inexpensive and relatively easy to implement. However, both methods are most commonly detected by slab gel electrophoresis, which can be labor-intensive, time-consuming, and often the methods are unable to produce high sensitivity and specificity without the use of multiple analysis conditions. Here, we demonstrate the first blinded study using microchip electrophoresis (ME)-SSCP/HA. We demonstrate the ability of ME-SSCP/HA to detect with 98% sensitivity and specificity >100 samples from the p53 gene exons 5-9 in a blinded study in an analysis time of <10 min.
authors: Northwestern University, Department of Chemical Engineering, Evanston, IL, USA.
authors: Christa N
authors: CN
authors: Hestekin
authors: Jennifer S
authors: JS
authors: Lin
authors: Lionel
authors: L
authors: Senderowicz
authors: John P
authors: JP
authors: Jakupciak
authors: Catherine
authors: C
authors: O'Connell
authors: Alfred
authors: A
authors: Rademaker
authors: Annelise E
authors: AE
authors: Barron
mesh_terms: D004247:DNA / Q000032:analysis / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods; D046209:Electrophoresis, Microchip / Q000379:methods*; D016158:Genes, p53; D020180:Heteroduplex Analysis / Q000379:methods*; D006801:Humans; D009154:Mutation; D009369:Neoplasms / Q000235:genetics; D018807:Polymorphism, Single-Stranded Conformational; D012107:Research Design; D012680:Sensitivity and Specificity
chemical_list: D004247:DNA