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Highly Rapid Amplification-Free and Quantitative DNA Imaging Assay

Please always quote using this URN: urn:nbn:de:bvb:20-opus-130500
  • 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 ofThere 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.show moreshow less

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Metadaten
Author: Tobias Klamp, Marta Camps, Benjamin Nieto, Francesc Guasch, Rohan T. Ranasinghe, Jens Wiedemann, Zdeněk Petrášek, Petra Schwille, David Klenerman, Markus Sauer
URN:urn:nbn:de:bvb:20-opus-130500
Document Type:Journal article
Faculties:Fakultät für Biologie / Theodor-Boveri-Institut für Biowissenschaften
Language:English
Parent Title (English):Scientific Reports
Year of Completion:2013
Volume:3
Issue:1852
Source:Scientific Reports 3, 1852; DOI:10.1038/srep01852 (2013)
DOI:https://doi.org/10.1038/srep01852
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Tag:assay systems; diseases; infectious diseases; laboratory techniques and procedures
Release Date:2016/07/07
EU-Project number / Contract (GA) number:216027
OpenAIRE:OpenAIRE
Licence (German):License LogoCC BY-NC-ND: Creative-Commons-Lizenz: Namensnennung, Nicht kommerziell, Keine Bearbeitung