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Digital PCR
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DOI：10.1073/pnas.96.16.9236.png)
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Abstract
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The identification of predefined mutations expected to be present in a minor fraction of a cell population is important for a variety of basic research and clinical applications. Here, we describe an approach for transforming the exponential, analog nature of the PCR into a linear, digital signal suitable for this purpose. Single molecules are isolated by dilution and individually amplified by PCR; each product is then analyzed separately for the presence of mutations by using fluorescent probes. The feasibility of the approach is demonstrated through the detection of a mutant ras oncogene in the stool of patients with colorectal cancer. The process provides a reliable and quantitative measure of the proportion of variant sequences within a DNA sample.
Keywords:
POLYMERASE CHAIN-REACTION
RAS GENE-MUTATIONS
MALIGNANT-MELANOMA
ONCOGENE MUTATIONS
TUMOR-PROGRESSION
MOLECULAR BEACONS
PERIPHERAL-BLOOD
POINT MUTATIONS
CANCER PATIENTS
GENOMIC DNA
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IF:
9.1
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10.8W
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73.5W
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