NDLI: Electronic detection of nucleic acids: a versatile platform for molecular diagnostics.

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Electronic detection of nucleic acids: a versatile platform for molecular diagnostics.

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Electronic detection of nucleic acids: a versatile platform for molecular diagnostics.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Umek, R. M. Lin, S. W. Vielmetter, J. Terbrueggen, R. H. Irvine, B. Yu, C. J. Kayyem, J. F. Yowanto, H. Blackburn, G. F. Farkas, D. H. Chen, Y. P.
Description	Country affiliation: United States Author Affiliation: Umek RM ( Clinical Micro Sensors Division of Motorola, Inc., Pasadena, California 91105, USA.)
Abstract	A novel platform for the electronic detection of nucleic acids on microarrays is introduced and shown to perform well as a selective detection system for applications in molecular diagnostics. A gold electrode in a printed circuit board is coated with a self-assembled monolayer (SAM) containing DNA capture probes. Unlabeled nucleic acid targets are immobilized on the surface of the SAM through sequence-specific hybridization with the DNA capture probe. A separate signaling probe, containing ferrocene-modified nucleotides and complementary to the target in the region adjoining the capture probe binding site, is held in close proximity to the SAM in a sandwich complex. The SAM allows electron transfer between the immobilized ferrocenes and the gold, while insulating the electrode from soluble redox species, including unbound signaling probes. Here, we demonstrate sequence-specific detection of amplicons after simple dilution of the reaction product into hybridization buffer. In addition, single nucleotide polymorphism discrimination is shown. A genotyping chip for the C282Y single nucleotide polymorphism associated with hereditary hemochromatosis is used to confirm the genotype of six patients' DNA. In addition, a gene expression-monitoring chip is described that surveys five genes that are differentially regulated in the cellular apoptosis response. Finally, custom modification of individual electrodes through sequence-specific hybridization demonstrates the potential of this system for infectious disease diagnostics. The versatility of the electronic detection platform makes it suitable for multiple applications in diagnostics and pharmacogenetics.
File Format	HTM / HTML
ISSN	15251578
e-ISSN	19437811
Journal	The Journal of Molecular Diagnostics
Issue Number	2
Volume Number	3
Language	English
Publisher	Elsevier
Publisher Date	2001-05-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Molecular Biology Nucleic Acids Chemistry Ultrastructure Oligonucleotide Array Sequence Analysis Instrumentation Apoptosis Base Pair Mismatch Dna, Complementary Metabolism Electrochemistry Genotype Molecular Sequence Data Polymorphism, Single Nucleotide Rna-directed Dna Polymerase Reverse Transcriptase Polymerase Chain Reaction Temperature
Content Type	Text
Resource Type	Article
Subject	Molecular Medicine Pathology and Forensic Medicine

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