Indicator Bacteriophage Detection for Rapid Microorganism Screening
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting microorganisms, such as bacteria, in biological, food, and water samples are time-consuming, often requiring several days due to the need for enrichment cultures and amplification steps, which is unsuitable for rapid identification of pathogens, especially in the context of food contamination and antibiotic-resistant bacteria.
Innovation Solution
Development of recombinant reproduction-deficient bacteriophages with an indicator gene in a late gene region, capable of specifically infecting target microorganisms, allowing for rapid detection of as few as 10-1 CFU/mL in samples without the need for enrichment cultures, using luciferase expression as a detection signal.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional microbiological tests with enrichment cultures are used, then sensitivity for detecting microorganisms is improved, but detection time increases to several days
Solution Approach 1:
The detection system performs preliminary actions by using phage infection to amplify the detectable signal from target microorganisms before final detection. The phage infects the bacteria and produces indicator proteins that can be detected at much lower concentrations than the original bacterial cells, effectively performing signal amplification in advance of the measurement step.
Solution Approach 2:
The patent introduces bacteriophage as an intermediary agent between the target microorganism and the detection system. The phage serves as a mediator that binds to specific bacterial surface receptors, enters the cell, and produces indicator proteins that reveal the presence of the target organism. This intermediary approach enables detection without requiring direct observation of the bacteria themselves.
2Measurement precision
If PCR tests with amplification steps are used, then sensitivity and selectivity are improved, but sample size requirements are limited and purification steps are needed
Solution Approach 1:
The patent extracts the amplification function from complex molecular biology procedures and transfers it to a biological system. Instead of using PCR to amplify DNA, the system uses phage infection to amplify the presence of the target organism through production of indicator proteins. This extraction of the amplification function simplifies the overall system by eliminating the need for DNA extraction, purification, and amplification steps.
Solution Approach 2:
The detection system uses the target microorganism's own biological machinery to perform the amplification function. The bacterium's cellular processes are harnessed to produce phage proteins that serve as detection signals. This self-service approach eliminates the need for external amplification equipment and complex reagent systems required by PCR.
3Measurement precision
If overnight enrichment incubation is used, then adequate sensitivity is achieved, but same-day results cannot be delivered
Solution Approach 1:
The patent changes the detection parameter from requiring large numbers of bacterial cells (achieved through overnight enrichment) to detecting phage-induced indicator proteins. By shifting the detection target from the bacteria themselves to the phage products they generate, the system achieves high sensitivity with much shorter incubation times, enabling same-day results while maintaining adequate 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
Enables rapid and sensitive detection of microorganisms like E. coli and Salmonella in as little as 2 hours, with high specificity and sensitivity, reducing the time required for detection to a fraction of traditional methods, and facilitating same-day results.
Implementation Method 1
phages bind to specific bacteria by recognizing and binding to specific surface receptors
Implementation Method 2
expression of the indicator gene following infection of the microorganism of interest results in production of an indicator gene product
Data Source
AI summary
Disclosed herein are compositions, methods, kits and systems for rapid detection of microorganisms using a reproduction-deficient indicator bacteriophage. The specificity of such reproduction-deficient indicator bacteriophage for binding and infecting particular microorganisms of interest allows targeted and sensitive detection of a microorganism of interest.