Digital quantification using amplified single-molecule detection

• DOI: 10.1038/nmeth916
• Publication Date: 2006-08-23T18:20:37Z
• Authors: Jonas Jarvius; Jonas Melin; Jenny Göransson; Johan Stenberg; Simon Fredriksson; Carlos Gonzalez-Rey; Stefan Bertilsson; Mats Nilsson
• Keywords: DNA, Bacterial; 0301 basic medicine; 03 medical and health sciences; Spectrometry, Fluorescence; Molecular Probe Techniques; Nanotechnology; DNA, Circular; Sensitivity and Specificity; Vibrio cholerae

We describe a scheme for biomolecule enumeration by converting nanometer-scale specific molecular recognition events mediated by rolling-circle amplification to fluorescent micrometer-sized DNA molecules amenable to discrete optical detection. Our amplified single-molecule detection (SMD) approach preserves the discrete nature of the molecular population, allowing multiplex detection and highly precise quantification of molecules over a dynamic range of seven orders of magnitude. We apply the method for sensitive detection and quantification of the bacterial pathogen Vibrio cholerae.