Antigen Detection Reference Zone Normalization
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Conventional antigen detection methods face challenges in reproducibility and reliability due to variations in sample concentration and reaction temperature, particularly in on-site diagnostics where skilled technicians are not always available.
Innovation Solution
A novel method where the same detection antibodies are bound to both a test zone and a reference zone using a reference substance with the same epitope as the antigen, allowing for controlled sample flow and reaction time, thereby improving sensitivity and minimizing the impact of sample concentration and temperature on detection accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional immunoassay methods are used for antigen detection, then the detection can be performed using standard laboratory equipment and procedures, but the measurement results vary depending on sample concentration and reaction temperature, reducing reliability
Solution Approach 1:
The invention changes the reference substance from a non-specific control to one containing the same epitope as the antigen. This parameter change ensures that the reference zone responds to the same binding conditions as the test zone, making the ratio calculation insensitive to variations in sample concentration and reaction temperature, thereby improving both reliability and measurement precision
Solution Approach 2:
The invention uses the reference zone signal as feedback to normalize the test zone signal. By calculating the ratio of test zone to reference zone signals, the system automatically compensates for variations in sample concentration and reaction conditions, improving measurement reliability without requiring manual intervention or specialized technician skills
2Reliability
If different detection antibodies are bound to test zone and reference zone to detect antigen specifically binding to test zone, then specific detection is achieved, but reproducibility and reliability are poor due to concentration and temperature variations
Solution Approach 1:
The invention uses the same detection antibody bound to both test zone and reference zone, creating homogeneity in the detection system. This eliminates variability introduced by using different antibodies with potentially different affinities and specificities, improving reproducibility while simplifying the system by reusing the same reagent
Solution Approach 2:
The detection antibody serves a dual function by being bound to both the test zone (for antigen detection) and the reference zone (for normalization). This multi-functionality improves reproducibility by ensuring consistent detection conditions while reducing the number of different reagents needed
3Productivity
If on-site diagnosis is performed without skilled technicians, then prompt diagnosis is enabled, but the impact of sample concentration and temperature on detection accuracy increases
Solution Approach 1:
The assay system performs self-normalization using the reference zone containing the same epitope. The ratio calculation automatically compensates for variations in sample concentration and temperature without requiring technician intervention or calibration, enabling accurate on-site diagnosis by unskilled personnel while maintaining measurement precision
Solution Approach 2:
The reference zone acts as an intermediary that mediates the effect of sample concentration and temperature variations. By including the same epitope in the reference zone, it serves as an internal control that captures these variations, allowing the ratio calculation to eliminate their impact on measurement accuracy
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 stability, reliability, and reproducibility of antigen detection, enabling accurate on-site diagnosis without specialized skills and reducing the influence of sample concentration and temperature variations.
Implementation Method 1
contacting an analysis sample with a detection antibody with which a marker generating a detectable signal is combined and which specifically binds to the antigen
Implementation Method 2
contacting a capture antibody with the resultant product of step (a), the capture antibody specifically binding to an antigen to be detected
Implementation Method 3
contacting the detection antibody, with which the marker generating a detectable signal is combined, with a reference substance which is bound to a surface of a solid substrate and which includes an epitope to which the detection antibody specifically binds
Implementation Method 4
measuring signals generated from the markers of the resultant product of step (b) and the resultant product of step (c)
Data Source
AI summary
The present invention provides a method for detecting an antigen in an analysis sample, the method including: (a) contacting an analysis sample with a detection antibody with which a marker generating a detectable signal is combined and which specifically binds to the antigen; (b) contacting a capture antibody with the resultant product of step (a), the capture antibody specifically binding to an antigen to be detected; (c) contacting the detection antibody, with which the marker generating a detectable signal is combined, with a reference substance which is bound to a surface of a solid substrate and which includes an epitope to which the detection antibody specifically binds; (d) measuring signals generated from the markers of the resultant product of step (b) and the resultant product of step (c); and (e) analyzing the measured signals to determine the presence or absence and amount of the antigen in the analysis sample. The method for detecting an antigen of the present invention can control the flow and the reaction time of an analysis sample, thereby improving sensitivity and minimizing the influences by the concentration of the analysis sample or the temperature of the detection reaction, and thus improving stability, reliability, and reproducibility of data, when compared with the conventional method for detecting an antigen. Accordingly, the method and apparatus for detecting an antigen of the present invention can be easily operated without specialized skills, thereby instantly obtaining the presence or absence and amount of detection antigen in the analysis sample through on-site diagnosis.