Nature Methods
- 3, 725 - 727 (2006)
Published online: 23 August 2006; | doi:10.1038/nmeth916
Digital quantification using amplified single-molecule detectionJonas Jarvius1, 5, Jonas Melin1, 2, 5, Jenny Göransson1, Johan Stenberg1, Simon Fredriksson1, 4, Carlos Gonzalez-Rey1, 3, Stefan Bertilsson3 & Mats Nilsson11
Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala University, 751 85 Uppsala, Sweden. 2
Department of Engineering Sciences, A˚ngström Laboratory, Uppsala University, 751 21 Uppsala, Sweden. 3
Limnology/Department of Ecology & Evolution, BMC, Uppsala University, 751 23 Uppsala, Sweden. 4
Present address: Stanford Genome Technology Center, Stanford University, 318 Campus Drive, Stanford, California 94305, USA. 5
These authors contributed equally to this work.
Correspondence should be addressed to Mats Nilsson mats.nilsson@genpat.uu.se 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.
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