Pathogen Detection via Dual Temperature Incubation
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Solution Overview
Problem
Current methods for detecting pathogenic microorganisms, such as Salmonella, Listeria, and EHEC, require skilled technicians, specialized equipment, and take several days to produce results, struggling with specificity and false positives/negatives due to the heterogeneity and size of microorganisms.
Innovation Solution
A method involving dual temperature incubation to prevent and then induce the production of specific proteins in microorganisms, followed by detection using temperature-regulated protein-specific reagents, allowing for rapid and specific detection without specialized equipment or extensive training.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If traditional cultural methods are used for pathogen detection, then detection accuracy can be maintained, but detection time is extended to several days and specialized equipment is required
Solution Approach 1:
The method performs preliminary enrichment of the pathogen population in the sample before detection, allowing even low levels of contamination to be amplified to detectable levels. This preliminary growth phase ensures that sufficient target organisms are present for rapid detection while maintaining sensitivity for low-level contamination.
Solution Approach 2:
The invention changes the temperature parameter during the detection process, using temperature-dependent expression of virulence genes as a discriminatory feature. By incubating at specific temperatures that trigger expression of pathogen-specific virulence factors, the method achieves rapid differentiation between pathogenic and non-pathogenic organisms without requiring several days of cultivation.
2Ease of operation
If traditional detection methods are used, then comprehensive pathogen identification is achieved, but the methods require skilled technicians and specialized equipment
Solution Approach 1:
The method replaces complex mechanical and equipment-based detection systems with a biochemical assay that detects temperature-regulated virulence gene expression. Instead of requiring sophisticated instrumentation and skilled operators to interpret complex results, the invention uses antibody-based detection of specific protein markers that are automatically expressed under controlled temperature conditions, simplifying the operational requirements while maintaining identification accuracy.
3Quantity of substance
If standard incubation temperatures are used for pathogen growth, then optimal growth is achieved, but production of detection-targeted proteins is inhibited
Solution Approach 1:
The method employs periodic temperature variation to achieve two opposing objectives sequentially. First, the sample is incubated at a temperature optimal for pathogen growth to increase the quantity of target organisms. Then, the temperature is adjusted to a different value that triggers expression of the detection-targeted virulence proteins. This periodic temperature cycling allows both growth and protein production to be optimized at different stages of the same process.
Solution Approach 2:
The growth phase at the first temperature serves as a preliminary action that amplifies the pathogen population before the detection phase. By allowing robust growth first, the method ensures sufficient biomass is present to generate detectable levels of the temperature-regulated virulence proteins when the temperature is subsequently adjusted, solving the contradiction between growth and protein production.
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 reduces detection time, improves sensitivity, and enhances specificity, enabling rapid and accurate identification of pathogens in various matrices without requiring specialized skills or equipment.
Implementation Method 1
culturing the pathogen in the sample at a first temperature wherein the first temperature prevents production of at least one endogenous protein and wherein the first temperature allows for optimal growth of the pathogen
Implementation Method 2
culturing the pathogen in the sample at a second temperature wherein the second temperature allows production of the at least one endogenous protein
Implementation Method 3
contacting the sample with a detection reagent that specifically binds to the at least one protein; thereby detecting the pathogen in the sample
Data Source
AI summary
The present invention relates generally to methods for detecting and identifying microorganisms and, more particularly, to methods for detecting microorganisms in a sample by incubating the sample at two temperatures to facilitate increased detection of the organism.