Document Detail


Paper based Solid-Phase Nucleic Acid Hybridization Assay using Immobilized Quantum Dots as Donors in Fluorescence Resonance Energy Transfer.
MedLine Citation:
PMID:  23272728     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
A paper based solid-phase assay is presented for transduction of nucleic acid hybridization using immobilized quantum dots (QDs) as donors in fluorescence resonance energy transfer (FRET). The surface of paper was modified with imidazole groups to immobilize QD-probe oligonucleotide conjugates that were assembled in solution. Green-emitting QDs (gQDs) were FRET paired with Cy3 acceptor. Hybridization of Cy3-labeled oligonucleotide targets provided the proximity required for FRET sensitized emission from Cy3, which served as an analytical signal. The assay exhibited rapid transduction of nucleic acid hybridization within minutes. Without any amplification steps, the limit of detection of the assay was found to be 300 fmol with the upper limit of the dynamic range at 5 pmol. The implementation of glutathione coated QDs for the development of nucleic acid hybridization assay integrated on a paper-based platform exhibited excellent resistance to non-specific adsorption of oligonucleotides, and showed no reduction in the performance of the assay in the presence of large quantities of non-complementary DNA. The selectivity of nucleic acid hybridization was demonstrated by single nucleotide polymorphism (SNP) detection at a contrast ratio of 19 to 1. The reuse of paper over multiple cycles of hybridization and dehybridization was possible, with less than 20% reduction in the performance of the assay in 5 cycles. This work provides an important framework for the development of paper based solid-phase QD-FRET nucleic acid hybridization assays that make use of a ratiometric approach for detection and analysis.
Authors:
M Omair Noor; Anna Shahmuradyan; Ulrich J Krull
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-12-29
Journal Detail:
Title:  Analytical chemistry     Volume:  -     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-12-31     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
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