PCR Quantitation Automation via Threshold Cycle Computation
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Solution Overview
Problem
Conventional nucleic acid analysis methods, such as PCR, suffer from reduced accuracy and user-induced variability due to subjective interpretation and require significant user input, especially in high-throughput applications, necessitating an improved method for automation and quantitative accuracy.
Innovation Solution
A method involving a computing system that determines reference and target threshold cycles for nucleic acid quantification, allowing for automated sample input quantity calculation and display, thereby enhancing automation and reproducibility.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional PCR quantitation methods are used, then user input and interpretation are required, but this leads to reduced accuracy and user-induced variability
Solution Approach 1:
The system performs self-service by automatically determining threshold cycles and calculating input quantities without requiring user interpretation. The processor automatically compares threshold cycles between reference and target nucleic acids and computes the sample input quantity, eliminating the need for user input in the quantitation calculation process.
Solution Approach 2:
The patent replaces manual mechanical interpretation with automated computational processing. Instead of users manually analyzing PCR data and determining quantitation, a processor automatically performs threshold cycle determination and input quantity calculation, substituting human cognitive processes with computational algorithms.
2Extent of automation
If manual analysis methods are used, then flexibility in interpretation is maintained, but automation capabilities are reduced
Solution Approach 1:
The patent introduces a processor as an intermediary between the PCR reaction data and the final quantitation result. This intermediary automatically performs threshold cycle determination and input quantity calculation, serving as a mediator that translates raw PCR data into quantitative results without requiring direct user intervention in the calculation process.
Solution Approach 2:
The system changes the operational parameters from manual interpretation to automated computational processing. By implementing automated threshold cycle determination and input quantity calculation, the system transforms the quantitation process from a manual analytical task to an automated computational task, increasing automation while managing complexity through standardized algorithms.
3Productivity
If conventional quantitation methods are used, then processing speed is limited, but processing many samples simultaneously becomes feasible
Solution Approach 1:
The system performs preliminary actions by automatically establishing threshold cycles for reference and target nucleic acids before performing the actual quantitation calculation. This preliminary determination of threshold cycles enables rapid processing of multiple samples, as the automated system can quickly compare pre-determined threshold values and compute input quantities without manual intervention for each sample.
Data Source
AI summary
A method for quantifying nucleic acid is provided. The method includes determining a first reference threshold cycle for a first predetermined input quantity for a reference nucleic acid, determining a first target threshold cycle for the first predetermined input quantity for a target nucleic acid, determining a second reference threshold cycle for a second predetermined input quantity for the reference nucleic acid, and determining a second target threshold cycle, by the processor, for the second predetermined input quantity for the target nucleic acid. The method further includes receiving a sample threshold cycle, determining a sample input quantity based on the first and second reference threshold cycle and the first and second target threshold cycle, and displaying the sample input quantity to a user.