Lysis Agent for Coliform Bacteria Detection in Milk
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Conventional methods for detecting coliform bacteria in milk, such as cultivation-based methods and immunological measurement methods, face challenges in achieving efficient lysis due to high protein and fat content in milk, leading to inadequate detection sensitivity and speed.
Innovation Solution
A method involving a lysis agent containing a lytic enzyme, specifically lysozyme, combined with anionic and nonionic surfactants, particularly sodium laurylsulfate and polyoxyethylene alkyl phenyl ether, is used to efficiently lyse coliform bacteria in milk, enhancing detection sensitivity and speed through improved antigen release.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If cultivation-based methods are used to detect coliform bacteria in milk, then detection reliability is improved, but detection time increases significantly (several days required)
Solution Approach 1:
The detection process is segmented into two distinct parts: (1) a rapid immunochromatographic screening test that provides preliminary results in minutes, and (2) a cultivation-based confirmation test for positive cases. This segmentation allows most negative cases to be resolved quickly while maintaining high reliability through confirmatory testing of positive results.
Solution Approach 2:
The immunochromatographic test performs preliminary detection of coliform bacteria using antigen-antibody reactions before committing to time-consuming cultivation procedures. This preliminary action filters out negative cases rapidly, reserving cultivation resources only for samples that require further confirmation.
2Speed
If immunochromatographic method is used for quick detection, then detection speed is improved, but detection sensitivity deteriorates due to inadequate lysis in high protein and fat content milk
Solution Approach 1:
A milk treatment solution is introduced as an intermediary substance between the milk sample and the immunochromatographic test. This solution contains proteases that break down proteins and lipases that break down fats in the milk, creating a treated sample that allows adequate antigen release and antibody binding while maintaining the rapid detection advantage of the immunochromatographic method.
3Reliability
If conventional lysis agents are used in milk samples, then lysis effectiveness is improved in simple media, but lysis efficiency deteriorates in high protein and fat content milk
Solution Approach 1:
The chemical composition parameters of the treatment solution are specifically optimized for milk matrices. The solution contains proteases (such as proteinase K) and lipases in controlled concentrations, along with detergents and buffers, creating a formulation that changes the chemical environment to enable effective lysis in high protein and fat content samples while maintaining compatibility with subsequent immunochromatographic detection.
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 quick, convenient, and highly sensitive detection of coliform bacteria in milk, enabling early-stage diagnosis and appropriate treatment of mastitis, particularly in dairy farms without the need for specialized equipment.
Implementation Method 1
a lysis agent containing a lytic enzyme, specifically lysozyme
Implementation Method 2
combined with anionic and nonionic surfactants, particularly sodium laurylsulfate and polyoxyethylene alkyl phenyl ether
Implementation Method 3
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, lysis treatment solution, detection method using an immunochromatographic device, and detection kit comprising an immunochromatographic device for detecting whether causative bacteria of mastitis are coliform bacteria or not by using milk of a livestock animal. There is provided a method for lysing coliform bacteria, which comprises the step of mixing a lysis agent containing a lytic enzyme, and at least one kind of anionic surfactant, and preferably further containing at least one kind of nonionic surfactant, with milk obtained form a livestock animal to lyse coliform bacteria existing in the milk. The lytic enzyme is preferably lysozyme.