PCR Baseline Fluorescence Analysis for High-Titer Detection
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Solution Overview
Problem
Conventional real-time PCR systems using sequence detection software (SDS) often fail to accurately detect high concentrations of viruses like parvovirus B19 in plasma samples, leading to incorrect identification and potential contamination of plasma resources.
Innovation Solution
A method that calculates the fluorescence signal during a baseline period and determines if it increases by a certain percentage, flagging samples as potentially high-titer, and includes software logic to recalculate and verify cycle threshold values to ensure accurate detection and prevent misinterpretation of high viral loads.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional sequence detection software is used to analyze PCR results, then the system operates with standard detection capabilities, but it fails to accurately detect high concentrations of viruses leading to false negatives
Solution Approach 1:
The patent applies preliminary action by performing baseline period analysis before final detection. The system calculates fluorescence signals during the baseline period (cycles 3-15) and compares them to later cycles to identify high-titer samples before they are misclassified. This preliminary fluorescence comparison prevents false negatives by flagging samples that show abnormal fluorescence patterns early in the amplification process.
2Ease of manufacture
If the system uses standard baseline correction methods, then processing is simple, but high-titer samples are misinterpreted as low or negative
Solution Approach 1:
The patent implements feedback by using the calculated fluorescence signal from the baseline period to adjust the interpretation of final PCR results. When the baseline fluorescence signal increases by a predetermined percentage or exceeds a threshold, the system flags the sample as high-titer and overrides the standard detection algorithm. This feedback mechanism ensures that samples with high viral loads are correctly identified despite atypical amplification patterns.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach enhances the accuracy of PCR results by correctly identifying high-titer samples, preventing false negatives, and ensuring proper sample identification, thereby reducing the risk of plasma resource contamination.
Implementation Method 1
Real-time polymerase chain reaction (PCR) systems are used for amplifying a sample, such as a DNA strand
Implementation Method 2
A fluorescence signal is calculated from PCR results provided during a baseline period
Data Source
AI summary
Methods of managing results of a real-time polymerase chain reaction (PCR) instrument and software associated with such methods are described herein. One disclosed method, among others, comprises calculating, from results of the real-time PCR instrument, a fluorescence signal of a sample during a cycle of a baseline period of the real-time PCR instrument. The method further comprises determining whether or not the fluorescence signal during the baseline period increases by at least a certain percentage compared to cycles outside the baseline period. The sample is flagged as a potentially high-titer sample when the fluorescence signal increases by at least the certain percentage.