A reliable and sensitive bead-based fluorescence assay for identification of nucleic acid sequences

Tobias Klamp; Idir Yahiatène; André Lampe; Mark Schüttpelz; Markus Sauer

doi:10.1117/12.874567

23 February 2011 A reliable and sensitive bead-based fluorescence assay for identification of nucleic acid sequences

Tobias Klamp, Idir Yahiatène, André Lampe, Mark Schüttpelz, Markus Sauer

Proceedings Volume 7905, Single Molecule Spectroscopy and Imaging IV; 79050N (2011) https://doi.org/10.1117/12.874567
Event: SPIE BiOS, 2011, San Francisco, California, United States

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.

Citation Download Citation

Tobias Klamp, Idir Yahiatène, André Lampe, Mark Schüttpelz, and Markus Sauer "A reliable and sensitive bead-based fluorescence assay for identification of nucleic acid sequences", Proc. SPIE 7905, Single Molecule Spectroscopy and Imaging IV, 79050N (23 February 2011); https://doi.org/10.1117/12.874567

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