Multi-test analysis of real-time nucleic acid amplification
First Claim
1. A method of determining the presence of a nucleic acid in a sample comprising the steps ofproviding a fluorescent entity capable of indicating the presence of the nucleic acid and capable of providing a signal related to the quantity of the nucleic acid, amplifying the nucleic acid through a plurality of amplification cycles in the presence of the fluorescent entity, measuring fluorescence intensity of the fluorescent entity at each of the plurality of amplification cycles to produce a fluorescent value for each cycle related to the quantity of the nucleic acid present at each cycle, obtaining an individual score from each of a plurality of tests performed on the fluorescent values obtained during the plurality of amplification cycles, the plurality of tests comprising a Confidence Interval Test and a Signal-to-Noise Ratio Test, and using the scores to ascertain whether the nucleic acid is present in the sample.
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Abstract
A method and device are described for analyzing a sample for the presence of an analyte wherein the analyte is contacted with a substrate to effect a measurable change selected from the group consisting of the quantity of the analyte, the quantity of the substrate, and the quantity of an optical or physical change to the substrate, wherein the analyte is contacted with the substrate for a predetermined time period, to generate a signal related to the measurable change. Scores are obtained from various tests performed on the signal data, and the scores are used to determine whether the substrate is present in the sample.
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Citations
20 Claims
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1. A method of determining the presence of a nucleic acid in a sample comprising the steps of
providing a fluorescent entity capable of indicating the presence of the nucleic acid and capable of providing a signal related to the quantity of the nucleic acid, amplifying the nucleic acid through a plurality of amplification cycles in the presence of the fluorescent entity, measuring fluorescence intensity of the fluorescent entity at each of the plurality of amplification cycles to produce a fluorescent value for each cycle related to the quantity of the nucleic acid present at each cycle, obtaining an individual score from each of a plurality of tests performed on the fluorescent values obtained during the plurality of amplification cycles, the plurality of tests comprising a Confidence Interval Test and a Signal-to-Noise Ratio Test, and using the scores to ascertain whether the nucleic acid is present in the sample.
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17. A method of determining the presence of a nucleic acid in a sample comprising the steps of
providing a fluorescent entity capable of indicating the presence of a nucleic acid and capable of providing a signal related to the quantity of the nucleic acid, amplifying the nucleic acid through a plurality of PCR amplification cycles in the presence of the fluorescent entity, measuring fluorescence intensity of the fluorescent entity at each of the plurality of amplification cycles to produce a fluorescent value for each cycle related to the quantity of the nucleic acid present at each cycle, obtaining a score from each of a plurality of tests performed on the fluorescent values obtained during the plurality of amplification cycles, each of the plurality of tests using the fluorescence values to generate the score, wherein said plurality of tests are selected from the group consisting of Confidence Interval Test, Signal-to-Noise Ratio Test, Channel Consistency Test, Efficiency Test, Function Ordering Test, Maximum to Baseline Comparison Test, and Late rise Test, and using the scores to ascertain whether the nucleic acid is present in the sample.
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18. A method for determining the presence of an analyte in the sample, comprising the steps of
contacting the analyte with a substrate to effect a measurable change selected from the group consisting of the quantity of the analyte, the quantity of the substrate, and the quantity of an optical or physical change to the substrate, wherein the analyte is contacted with the substrate for a predetermined time period, generating a signal related to the measurable change, measuring signal intensity during said predetermined time period, such that intensity values are obtained for a plurality of time points, obtaining an individual score from each of a plurality of tests performed on the intensity values, the plurality of tests comprising a confidence interval test and a signal to noise ratio test, and using the scores to ascertain whether the analyte is present in the sample.
Specification