Nucleic Acid Analyzer Calibration Using a Single Adjuster
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Solution Overview
Problem
Existing methods for quantifying nucleic acids in real-time amplification procedures face challenges in achieving accurate quantitation across a wide dynamic range, particularly when using a single calibration standard, due to variations in reaction rates and primer sequences, and require multiple calibration standards or stored calibration curves, which can be costly and inefficient.
Innovation Solution
A method involving the use of dual calibration curves and a pivot point to establish an adjusted calibration curve on a local instrument, allowing for accurate quantitation using a single calibration standard by normalizing amplification indicia and maintaining a constant relative relationship between the curves, even when performed on different instruments or with varying reaction conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple calibration standards are used to achieve accurate quantitation across a wide dynamic range, then measurement precision is improved, but device complexity and cost increase due to requiring multiple calibrators and stored calibration curves
Solution Approach 1:
The patent divides the calibration process into two segments: a master calibration curve prepared once with multiple standards, and a local calibration curve prepared quickly with a single standard. This segmentation allows comprehensive calibration data to be obtained without requiring multiple standards for each measurement, reducing complexity while maintaining precision.
Solution Approach 2:
The master calibration curve is prepared in advance as a preliminary action, storing comprehensive calibration data that can be reused. This preliminary calibration eliminates the need to repeatedly use multiple standards, reducing both device complexity and reagent consumption while maintaining quantitation accuracy.
2Measurement precision
If multiple calibration standards are used to establish accurate calibration curves, then measurement precision is improved, but loss of substance increases due to consumption of limited reagents purchased in kit form
Solution Approach 1:
The master calibration curve is prepared once in advance using all necessary calibration standards, consuming the reagents in a single preliminary action. Subsequent measurements only require a single calibration standard, dramatically reducing ongoing reagent consumption and preventing loss of limited kit-form reagents.
Solution Approach 2:
The patent recovers the value of calibration standards by using them once to create a reusable master calibration curve. Instead of consuming multiple standards with each measurement, the calibrated data is stored and reused, effectively recovering the reagent investment while maintaining measurement precision.
3Ease of operation
If a single calibration standard is used to simplify workflows and reduce costs, then ease of operation is improved, but measurement precision deteriorates due to inability to accurately quantitate across wide dynamic range
Solution Approach 1:
The master calibration curve serves as an intermediary that bridges the gap between using a single calibration standard and achieving accurate quantitation across a wide dynamic range. It mediates by providing pre-established calibration data that compensates for the limitations of using only one standard, maintaining precision while simplifying operation.
Solution Approach 2:
The patent creates a copy of comprehensive calibration data in the form of a master calibration curve that can be reused indefinitely. This copied calibration information allows a single calibration standard to achieve the same quantitation accuracy that would otherwise require multiple standards, improving ease of operation without sacrificing precision.
4Productivity
If stored calibration curves are used to avoid running multiple calibrators each time, then productivity is improved, but device complexity increases due to requiring preparation and storage of full calibration plots
Solution Approach 1:
The calibration system is segmented into a comprehensive master calibration curve prepared once and a simplified local calibration curve prepared quickly. This segmentation allows the system to benefit from both approaches: the thoroughness of multiple standards initially, and the speed of single-standard calibration subsequently, improving productivity without excessive complexity.
Solution Approach 2:
The master calibration curve is prepared as a preliminary action, storing comprehensive calibration data that enables rapid subsequent calibrations. This preliminary work eliminates the need to repeatedly manage and store multiple full calibration plots, improving productivity while reducing the ongoing complexity of calibration data management.
Data Source
AI summary
Method and system for quantifying target nucleic acids using real-time amplification and internal calibration adjustment. The invention employs dual reference calibration curves for approximating a complete calibration curve from only a single adjustment calibrator amplified on the instrument that is to be calibrated.