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Digital quantification using amplified single-molecule detection
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DOI：10.1038/NMETH916.png)
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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.
Keyword:
ROLLING-CIRCLE AMPLIFICATION
PADLOCK PROBES
DNA-MOLECULES
REPLICATION
PCR
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