Streptococcus Lysis Agent for Milk Detection
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Solution Overview
Problem
Conventional methods for detecting Streptococcus bacteria in milk, such as cultivation-based methods, are slow and unsuitable for quick identification, while immunological methods face challenges in efficiently lysing bacterial cells due to high protein and fat content in milk, leading to inadequate antigen release for detection.
Innovation Solution
A lysis method using a lysis agent containing lysozyme, labiase, and β-N-acetylglucosaminidase, along with a nonionic surfactant, to efficiently lyse Streptococcus bacteria in milk, allowing for the release of specific antigens for sensitive detection using immunochromatographic methods.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If cultivation-based methods are used to detect Streptococcus bacteria in milk, then detection accuracy is improved, but detection time increases significantly
Solution Approach 1:
The patent applies preliminary action by pre-coating the test line on the immunochromatographic strip with antibodies specific to Streptococcus bacteria. This preparation is done in advance during manufacturing, so that when milk samples are tested, the detection can proceed immediately without requiring time-consuming cultivation steps. The pre-prepared antibodies are ready to bind to bacterial antigens directly in the milk sample.
Solution Approach 2:
The patent replaces the mechanical/cultivation-based detection system with an immunological detection system. Instead of using culture media and incubation procedures (mechanical/biological cultivation), the invention uses antibody-antigen binding reactions on a solid-phase immunochromatographic strip. This substitution enables rapid detection within minutes while maintaining high accuracy through specific immunological recognition.
2Speed
If immunochromatographic method is used for quick detection, then detection speed is improved, but lysis efficiency of bacterial cells deteriorates due to high protein and fat content in milk
Solution Approach 1:
The patent applies parameter changes by modifying the chemical composition of the lysis buffer to overcome the inhibitory effects of milk proteins and fats. The lysis buffer contains specific detergents, salts, and enzymes that are optimized to disrupt bacterial cell walls effectively in the presence of milk matrix components. By adjusting parameters such as pH, ionic strength, and detergent concentration, the buffer achieves reliable cell lysis despite the challenging sample matrix.
Solution Approach 2:
The patent uses a composite lysis buffer system that combines multiple components with complementary functions. The buffer includes detergents for membrane disruption, enzymes for cell wall degradation, and protective agents that prevent protein aggregation. This composite formulation works synergistically to achieve effective cell lysis in milk samples, overcoming the limitations of single-component lysis systems.
3Ease of operation
If conventional lysis methods are used on milk samples, then simplicity of operation is improved, but antigen release from bacterial cells deteriorates
Solution Approach 1:
The patent applies universality by designing a lysis buffer that performs multiple functions simultaneously. The buffer not only lyses bacterial cells but also protects released antigens from degradation, prevents protein aggregation, and maintains antigen stability throughout the detection process. This multi-functional buffer simplifies the overall procedure by eliminating the need for separate steps to protect or stabilize antigens after lysis.
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
The method enables highly efficient lysis of Streptococcus bacteria, facilitating rapid and accurate detection of Streptococcus uberis and Streptococcus agalactiae, thereby supporting early therapeutic intervention for mastitis.
Implementation Method 1
a lysis method using a lysis agent containing lysozyme, labiase, and β-N-acetylglucosaminidase, to efficiently lyse Streptococcus bacteria in milk
Implementation Method 2
lysis agent containing lysozyme, labiase, and β-N-acetylglucosaminidase
Implementation Method 3
A lysis method using a lysis agent containing lysozyme, labiase, and β-N-acetylglucosaminidase, along with a nonionic surfactant
Implementation Method 4
identification methods based on an antigen-antibody reaction using an antibody directed to an ingredient specific to a causative bacterium
Data Source
AI summary
The object is to provide a lysis method and lysis treatment solution for efficiently lysing cells of various Streptococcus bacteria in milk of a livestock animal to release a specific antigen substance contained in the cells for detecting whether causative bacterium of mastitis is a Streptococcus bacterium or not by using the milk, as well as a detection method using an immunochromatographic device. There is provided a method for lysing a Streptococcus bacterium, which comprises the step of mixing a lysis agent containing a lytic enzyme with milk obtained form a livestock animal to lyse a Streptococcus bacterium existing in the milk. The lytic enzyme is preferably at least one selected from the group consisting of lysozyme, labiase, and β-N-acetylglucosaminidase.