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Proceedings Paper

A reliable and sensitive bead-based fluorescence assay for identification of nucleic acid sequences
Author(s): Tobias Klamp; Idir Yahiatène; André Lampe; Mark Schüttpelz; Markus Sauer
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Paper Abstract

The sensitive and rapid detection of pathogenic DNA is of tremendous importance in the field of diagnostics. We demonstrate the ability of detecting and quantifying single- and double-stranded pathogenic DNA with picomolar sensitivity in a bead-based fluorescence assay. Selecting appropriate capturing and detection sequences enables rapid (2 h) and reliable DNA quantification. We show that synthetic sequences of S. pneumoniae and M. luteus can be quantified in very small sample volumes (20 μL) across a linear detection range over four orders of magnitude from 1 nM to 1 pM, using a miniaturized wide-field fluorescence microscope without amplification steps. The method offers single molecule detection sensitivity without using complex setups and thus volunteers as simple, robust, and reliable method for the sensitive detection of DNA and RNA sequences.

Paper Details

Date Published: 23 February 2011
PDF: 7 pages
Proc. SPIE 7905, Single Molecule Spectroscopy and Imaging IV, 79050N (23 February 2011); doi: 10.1117/12.874567
Show Author Affiliations
Tobias Klamp, Julius-Maximilians-Univ. Würzburg (Germany)
Idir Yahiatène, Bielefeld Univ. (Germany)
André Lampe, Bielefeld Univ. (Germany)
Mark Schüttpelz, Bielefeld Univ. (Germany)
Markus Sauer, Julius-Maximilians-Univ. Würzburg (Germany)

Published in SPIE Proceedings Vol. 7905:
Single Molecule Spectroscopy and Imaging IV
Jörg Enderlein; Zygmunt Karol Gryczynski; Rainer Erdmann, Editor(s)

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