Periodontopathic Bacteria Detection Using Reducing Agents
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Solution Overview
Problem
Current methods for detecting periodontopathic bacteria, particularly those with trypsin-like enzymatic activity, require special equipment and optimal temperature conditions, limiting their sensitivity and speed, especially for rapid screening at room temperature.
Innovation Solution
The method involves using a compound with antioxidant effects or one that protects SH groups and cleaves disulfide bonds, such as L-ascorbic acid, L-cysteine hydrochloride, glutathione, DTT, thioglycolic acid, thioglycerol, mercaptoethanol, or TCEP, to analyze trypsin-like enzymatic activity with high sensitivity at room temperature in less than 10 minutes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If trypsin-like enzymatic activity is measured at optimal temperature (50-60°C), then sensitivity of detection is improved, but special equipment capable of maintaining optimal temperature is required
Solution Approach 1:
The invention changes the temperature parameter from optimal 50-60°C to room temperature (20-25°C) by introducing a reducing agent that stabilizes enzyme activity at lower temperatures, eliminating the need for temperature control equipment while maintaining detection sensitivity
Solution Approach 2:
A reducing agent acts as an intermediary substance that enables the enzyme to function effectively at room temperature by preventing oxidation and maintaining enzyme structure, thereby bridging the gap between optimal temperature requirements and room temperature operation
2Ease of operation
If trypsin-like enzymatic activity is measured at room temperature without reducing agents, then no special equipment is needed, but sufficient sensitivity cannot be obtained
Solution Approach 1:
The reducing agent serves as a mediator that enables room temperature measurement by stabilizing the enzyme, allowing both operational simplicity and detection sensitivity to be achieved simultaneously
Solution Approach 2:
The invention modifies the chemical environment by introducing reducing agents that change the stability parameters of the enzyme at room temperature, enabling sensitive detection without temperature control equipment
3Measurement precision
If enzyme activity analysis is performed for 10 minutes at room temperature, then sensitivity is improved, but time constraint for patient is increased
Solution Approach 1:
The reducing agent changes the reaction kinetics parameters by stabilizing the enzyme-substrate complex, enabling sensitive detection to be achieved in less than 10 minutes at room temperature
Solution Approach 2:
The reducing agent maintains continuous enzyme activity at room temperature by preventing oxidation, allowing the reaction to proceed effectively throughout the shortened measurement period without losing sensitivity
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 allows for the detection of periodontopathic bacteria, specifically P.g, T.d, and T.f, with high sensitivity and in a shorter time frame than traditional methods, facilitating rapid screening and diagnosis of periodontal disease without the need for special equipment.
Implementation Method 1
a compound having an antioxidant effect or a compound having action of protecting SH groups (mercapto groups) and cleaving disulfide bonds is present when analyzing trypsin-like enzymatic activity
Implementation Method 2
analyze trypsin-like enzymatic activity with high sensitivity at room temperature in less than 10 minutes
Data Source
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AI summary
The present invention makes it possible to simply, rapidly, and with high sensitivity detect periodontopathic bacteria at room temperature and in under ten minutes (approximately 2-8 minutes). The arginine-specific peptidase activity of three strains of periodontopathic bacteria, i.e., Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia, are analyzed at room temperature, in under ten minutes, and with high sensitivity. A first compound that has an anti-oxidizing effect and/or a second compound that protects SH groups (mercapto groups) and has a disulfide bond-cleaving effect are present during analysis of a sample. The first compound is at least one among L-ascorbic acid, L-cysteine hydrochloride, and glutathione, and the second compound is at least one among DTT, thioglycolic acid, thioglycerol, mercaptoethanol, and TCEP.