Indicator Bacteriophage Detection for Same-Day Microorganism Testing
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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 traditional enrichment cultures and lack the ability to provide same-day results, while rapid methods like PCR are limited by sample size and require lengthy enrichment steps.
Innovation Solution
The use of recombinant reproduction-deficient bacteriophages with indicator genes inserted into late gene regions, which are engineered to specifically infect target microorganisms and express detectable products, allowing for rapid detection without culturing enrichment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional enrichment cultures are used for microorganism detection, then sensitivity is improved, but detection time increases to several days
Solution Approach 1:
The patent segments the detection process by using specific phage types that naturally bind to and concentrate target bacteria, eliminating the need for lengthy general enrichment cultures. The phage-mediated concentration step replaces days of bulk bacterial cultivation with a targeted, rapid binding process.
Solution Approach 2:
The patent introduces bacteriophages as intermediary agents that mediate between the sample and detection system. These phages specifically infect target bacteria, amplifying the detectable signal while reducing the time required for bacterial enrichment through their natural host-specific replication cycle.
2Loss of time
If rapid detection methods like PCR are used, then detection time is reduced, but sample size requirements limit sensitivity
Solution Approach 1:
The patent performs preliminary concentration of target bacteria using phage binding before the actual detection step. This pre-concentration ensures that even small sample volumes contain sufficient target organisms for sensitive detection, eliminating the need for large sample sizes required by conventional PCR methods.
3Measurement precision
If overnight enrichment is performed to achieve adequate sensitivity, then detection sensitivity is improved, but same-day results cannot be delivered
Solution Approach 1:
The patent changes the key parameter from bulk bacterial growth rate to phage-binding kinetics. Since phage-bacteria binding occurs rapidly regardless of bacterial density, the method achieves high sensitivity without requiring overnight enrichment, enabling same-day results while maintaining detection 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 enables rapid and sensitive detection of microorganisms, including as few as a single bacterium, with high specificity and accuracy, reducing the time required for detection to hours rather than days.
Implementation Method 1
incubating the sample with a reproduction-deficient indicator phage that infects the microorganism of interest
Implementation Method 2
such that expression of the indicator gene following infection of the microorganism of interest results in production of an indicator gene product
Implementation Method 3
detecting the indicator gene product, wherein positive detection of the indicator gene product indicates that the microorganism of interest is present in the sample
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.