Nucleic Acid Quantitation Calibration Using Single Fixed Point
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Solution Overview
Problem
Current methods for quantifying nucleic acids using real-time amplification procedures face challenges in achieving accurate and simplified internal calibration, particularly when using a single calibration standard across a wide dynamic range, and are costly due to the need for multiple calibration standards and reagents.
Innovation Solution
A method involving the establishment of an adjusted calibration curve using a fixed-point and a single internal calibrator, where the fixed-point is determined using a different instrument or kit, allowing for co-amplification of the analyte and calibrator to normalize amplification indicia, enabling accurate quantitation with minimal calibration standards and reagents.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple calibration standards are used for internal calibration adjustment, then measurement precision is improved, but device complexity and reagent consumption increase
Solution Approach 1:
The patent extracts only the essential calibration information by using a single calibration standard to determine a calibration factor, rather than requiring multiple standards. This simplifies the calibration process while maintaining quantitation accuracy across the dynamic range.
Solution Approach 2:
The patent changes the calibration approach from using multiple concentration points to using a single calibration standard with a determined calibration factor. This parameter change simplifies the calibration process while the mathematical model maintains accuracy across the full dynamic range.
2Measurement precision
If multiple calibration standards are used, then measurement precision is improved, but loss of substance increases due to reagent consumption
Solution Approach 1:
The patent extracts only the essential calibration information from a single calibration standard, avoiding the need to consume multiple calibration standards and their associated reagents. This reduces reagent waste while maintaining quantitation accuracy.
Solution Approach 2:
The patent uses a single calibration standard that can be depleted after one use, replacing the need for multiple long-lived calibration standards. This approach reduces overall reagent consumption while maintaining measurement precision.
3Device complexity
If a single calibration standard is used, then device complexity is reduced, but measurement precision deteriorates across wide dynamic range
Solution Approach 1:
The patent introduces a calibration factor determined from a single standard and applies it across the full dynamic range through mathematical modeling. This allows one standard to accurately calibrate measurements across multiple orders of magnitude.
Solution Approach 2:
The calibration factor determined from a single calibration standard serves multiple functions: it calibrates the instrument response, establishes the dynamic range, and enables accurate quantitation across the full measurement range. This multi-functionality maintains precision while simplifying the process.
4Measurement precision
If conventional calibration methods are used, then measurement precision is maintained, but loss of time increases due to repeated calibration procedures
Solution Approach 1:
The patent performs calibration using a single standard that can be stored and reused, eliminating the need for repeated time-consuming calibration procedures. The calibration factor is determined once and applied to subsequent measurements, saving time while maintaining accuracy.
Solution Approach 2:
The calibration factor established from a single standard continues to be valid for multiple measurement sessions, allowing continuous useful action without interruption for recalibration. This maintains measurement precision while eliminating repeated calibration time losses.
Data Source
AI summary
Method and system for quantifying target nucleic acids using real-time amplification and internal calibration adjustment. The approach employs a single fixed data point in combination with a single adjustment calibrator amplified on the instrument that is to be calibrated for approximating a complete calibration curve.