Automatic threshold setting and baseline determination for real-time PCR
AssigneeApplera Corporation
InventorsDavid Woo, Clinton Lewis, Nasser Abbasi
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Abstract
The invention discloses a system and methods for quantitating the presence of nucleic acid sequences by evaluation of amplification data generated using real-time PCR. In one aspect, the methods may be adapted to identify a threshold and threshold cycle for one or more reactions based upon evaluation of exponential and baseline regions for each amplification reaction. The methodology used in the analysis may be readily automated such that subjective user interpretation of the data is substantially reduced or eliminated.
Claims12
A method of generating a baseline associated with one or more amplification reactions, comprising:
- receiving signal information including detected reporter label intensities generated by amplifying at least one target sequence in the presence of a detectable reporter construct;
- identifying a baseline region bounded by a lower bound of an exponential region of the signal information, the lower bound being generated by detecting incremental differences in the signal information starting at an upper bound of the exponential region;
- and
- generating a characteristic equation of the baseline region by performing a regression analysis of the detected intensities in the baseline region.
A system for generating a baseline associated with one or more amplification reactions, comprising:
- a data collection module that receives signal information including detected reporter label intensities generated by amplifying at least one target sequence in the presence of a detectable reporter construct;
- and
- a data processing module configured to
- identify a baseline region bounded by a lower bound of an exponential region of the signal information, the lower bound being generated by detecting incremental differences in the signal information starting at an upper bound of the exponential region, and
- generate a characteristic equation of the baseline region by performing a regression analysis of the detected intensities in the baseline region.
Dependencies are inferred from the claim text.