TY  - JOUR
A1  - Klamp, Tobias
A1  - Camps, Marta
A1  - Nieto, Benjamin
A1  - Guasch, Francesc
A1  - Ranasinghe, Rohan T.
A1  - Wiedemann, Jens
A1  - Petrášek, Zdeněk
A1  - Schwille, Petra
A1  - Klenerman, David
A1  - Sauer, Markus
T1  - Highly Rapid Amplification-Free and Quantitative DNA Imaging Assay
JF  - Scientific Reports
N2  - There is an urgent need for rapid and highly sensitive detection of pathogen-derivedDNAin a point-of-care (POC) device for diagnostics in hospitals and clinics. This device needs to work in a ‘sample-in-result-out’ mode with minimum number of steps so that it can be completely integrated into a cheap and simple instrument. We have developed a method that directly detects unamplified DNA, and demonstrate its sensitivity on realistically sized 5 kbp targetDNA fragments of Micrococcus luteus in small sample volumes of 20 mL. The assay consists of capturing and accumulating of target DNA on magnetic beads with specific capture oligonucleotides, hybridization of complementary fluorescently labeled detection oligonucleotides, and fluorescence imaging on a miniaturized wide-field fluorescence microscope. Our simple method delivers results in less than 20 minutes with a limit of detection (LOD) of,5 pMand a linear detection range spanning three orders of magnitude.
KW  - laboratory techniques and procedures
KW  - diseases
KW  - infectious diseases
KW  - assay systems
Y1  - 2013
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130500
VL  - 3
IS  - 1852
ER  -