Multi-Temperature PCR Signal Estimation for Target Analyte Separation
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Solution Overview
Problem
Existing methods for obtaining approximate signals for multiple target analytes fail to accurately reflect the approximate signals for each of a plurality of target analytes in real-time PCR due to limitations in reflecting detection environment differences and reliance on pre-determined reference values.
Innovation Solution
A method using a computer device to perform joint-estimation on approximate function parameters and magnitude parameters from signal generation reactions at different temperatures, allowing for the estimation of optimized signal values for each target analyte without relying on pre-determined reference values.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If reference values are obtained from other reaction wells in advance, then the detection process is simplified, but the detection accuracy is reduced due to inability to reflect detection environment differences
Solution Approach 1:
The system performs self-calibration by automatically extracting reference values from the current reaction well's data without requiring separate calibration wells. The processor identifies amplification curves, determines baseline regions, and calculates reference values directly from the sample being tested, eliminating the need for pre-obtained reference values from other wells while maintaining high accuracy.
Solution Approach 2:
The system performs preliminary processing of the signal data by automatically identifying amplification curves and baseline regions before extracting reference values. This preliminary action includes detecting the presence of amplification curves, determining baseline regions through automated algorithms, and preparing the data for accurate reference value extraction, all within the same reaction well.
2Ease of manufacture
If pre-determined reference values are used, then the method is easier to implement, but the signal distortion increases due to environmental differences
Solution Approach 1:
The system dynamically adjusts the reference value parameter based on the specific detection environment of each reaction well. Instead of using fixed pre-determined reference values, the processor calculates reference values that adapt to local environmental conditions such as detection device characteristics and reaction well variations, thereby maintaining signal accuracy across different conditions.
3Adaptability or versatility
If multiple target analytes are detected using different detection temperatures, then the detection capability is enhanced, but the signal separation difficulty increases
Solution Approach 1:
The system segments the detection process by performing separate joint-estimation for each target analyte based on their respective amplification curves at different detection temperatures. The processor identifies and processes amplification curves for first and second target analytes separately, extracting reference values and signals for each analyte independently, thereby simplifying the separation of overlapping signals from multiple analytes.
Data Source
AI summary
A method for obtaining an approximate signal for each of a plurality of target analytes performed by a computer device includes: obtaining, from a signal generation reaction for a first target analyte and a second target analyte in a sample, a first data set measured at a first detection temperature and a second data set measured at a second detection temperature; and by using the first and the second data set, performing a joint-estimation on (a) a value of a parameter of at least one among an approximate function for approximating signal values dependent on the presence of the first target analyte and an approximate function for approximating signal values dependent on the presence of the second target analyte; and (b) a value of a magnitude parameter indicating a relationship of signal values dependent on the presence of the first target analyte between the first and the second detection temperature.