Dual Indicator Biological Activity Detection Method
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Solution Overview
Problem
Existing methods for detecting biological activities in samples face challenges due to interference from high concentrations of indicator reagents, which hinder the detection of biological derivatives, particularly in biological sterilization processes where accurate and rapid detection is crucial.
Innovation Solution
The method employs a system with two indicator reagents, where a first indicator reagent with a specific absorbance spectrum is converted into a biological derivative, and a second indicator reagent, a fluorogenic enzyme substrate, is used to detect enzymatic activities, utilizing a substrate to concentrate the first indicator reagent and reduce interference, allowing for sensitive detection of the biological derivative.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a high concentration of first indicator reagent is used to detect biological activity, then the sensitivity of detection is improved, but the interference with the optical detection of the second indicator reagent increases
Solution Approach 1:
The detection system is divided into two separate detection channels: one for the first indicator reagent (detecting biological activity) and one for the second indicator reagent (fluorescence detection). This segmentation allows each indicator to operate at optimal concentrations without mutual interference, as they are detected through different optical paths and wavelengths.
Solution Approach 2:
The patent uses wavelength-specific optical filters as intermediaries between the light sources and detectors. These filters selectively transmit only the relevant wavelengths to each detector, blocking interference from the other indicator reagent. This allows the first indicator reagent to be present at high concentrations without affecting the fluorescence detection of the second indicator reagent.
2Measurement precision
If two indicator systems are used to detect specific cells, then the accuracy of cell identification is improved, but the complexity of the detection system increases
Solution Approach 1:
The detection device is designed with multi-functional capabilities: a single instrument performs both absorbance detection (for the first indicator reagent) and fluorescence detection (for the second indicator reagent). The device uses multiple light sources and detectors that can operate across different wavelength ranges, allowing one instrument to execute multiple detection functions simultaneously, thereby reducing overall system complexity.
Solution Approach 2:
The patent combines two indicator systems into a single reaction mixture, allowing both indicators to work together in the same sample. The detection device merges absorbance and fluorescence detection capabilities into one instrument, with integrated optical paths and control systems that coordinate both detection modes, simplifying the overall detection process while maintaining high identification 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 enables rapid and sensitive detection of biological activities by sequestering the interfering first indicator reagent, facilitating the observation of the biological derivative, thereby improving the accuracy and efficiency of biological activity detection in samples, including those subjected to sterilization processes.
Implementation Method 1
a substrate that receives and concentrates the first indicator reagent from an aqueous mixture
Implementation Method 2
a second indicator reagent that is converted by the first predetermined enzymatic activity to a second biological derivative with a second emission spectrum
Implementation Method 3
The methods include a second indicator system, such as 4-methylumbelliferyl-p-D-glucuronic acid, which is used to detect the enzyme β-glucuronidase
Implementation Method 4
a first indicator reagent with a first absorbance spectrum, wherein the first indicator reagent comprises a pH indicator dye or a redox dye, wherein the first indicator reagent can be converted by a first predetermined enzymatic activity to a first biological derivative
Implementation Method 5
detecting a presence or absence of fluorescence from the second biological derivative comprises using the instrument to detect the second biological derivative, wherein the optical path does not intersect any portion of the substrate
Data Source
Figure 1A~1C
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AI summary
The present invention provides method of detecting a predetermined biological activity. The method includes using an aqueous mixture comprising a first indicator reagent with a first absorption spectrum and a second indicator reagent. The second indicator reagent is converted by the predetermined biological activity to a second biological derivative with a second emission spectrum. The first absorbance spectrum includes detectable absorbance in at least a portion of wavelengths present in the second emission spectrum. The first indicator reagent is received and concentrated from an aqueous liquid by a substrate, facilitating the detection of the second biological derivative.