Multi-test analysis of real-time nucleic acid amplification
AssigneeUniversity of Utah Research Foundation
InventorsDavid J. Eyre, Carl T. Wittwer
Life of this patent
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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.
Claims33
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, 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.
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 a score from each of a plurality of tests, each of the plurality of tests using the fluorescence values to generate the score, and
- using the scores to ascertain whether the nucleic acid is present in the sample.
A device for determining the presence of a nucleic acid in a sample comprising
- an instrument for temperature cycling to amplify the nucleic acid,
- a fluorimeter for detecting fluorescence during amplification of the nucleic acid, the fluorescence obtained from a fluorescent entity capable of providing a signal related to the quantity of the nucleic acid, and
- a processor for performing analysis routines, wherein the processor is programmed to obtain a score from each of a plurality of tests, each of the plurality of tests using fluorescence values measured by the fluorimeter to generate the score, and to use the scores to ascertain whether the nucleic acid is present in the sample.
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, 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.
Dependencies are inferred from the claim text.