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Blinded study determination of high sensitivity and specificity microchip electrophoresis‐SSCP/HA to detect mutations in the p53 gene
Author(s) -
Hestekin Christa N.,
Lin Jennifer S.,
Senderowicz Lionel,
Jakupciak John P.,
O'Connell Catherine,
Rademaker Alfred,
Barron Annelise E.
Publication year - 2011
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201100396
Subject(s) - heteroduplex , single strand conformation polymorphism , microbiology and biotechnology , biology , gel electrophoresis , blinded study , computational biology , gene , electrophoresis , mutation , genetics , medicine
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.