Digital quantification using amplified single-molecule detection.
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.
Jarvius
Jonas
J
Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala University, 751 85 Uppsala, Sweden.
Melin
Jonas
J
Göransson
Jenny
J
Stenberg
Johan
J
Fredriksson
Simon
S
Gonzalez-Rey
Carlos
C
Bertilsson
Stefan
S
Nilsson
Mats
M
D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics*; D004270:DNA, Circular / Q000737:chemistry / Q000235:genetics*; D015336:Molecular Probe Techniques; D036103:Nanotechnology; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods; D014734:Vibrio cholerae / Q000235:genetics*
D004269:DNA, Bacterial; D004270:DNA, Circular