Recombinant Bacteriophage Detection for Same-Day Microorganism Testing
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Solution Overview
Problem
Current methods for detecting microorganisms, such as bacteria and viruses, are time-consuming, requiring several days due to traditional culturing and enrichment processes, which is unsuitable for rapid identification of pathogens in food, water, and clinical samples, especially with the rise of antibiotic-resistant strains.
Innovation Solution
The use of recombinant bacteriophages that are genetically modified to include a reporter gene, allowing for rapid detection of microorganisms by binding to them and generating an indicator moiety, which can be detected using a self-contained apparatus with separate compartments for sample incubation and substrate reaction, reducing the need for traditional enrichment and shortening the detection time to under 12 hours.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional microbiological tests with enrichment cultures are used, then sensitivity and selectivity are improved, but detection time increases to several days
Solution Approach 1:
The patent replaces traditional mechanical/biological enrichment culturing with a molecular biology approach using PCR amplification. The PCR process directly amplifies target DNA sequences from the sample without requiring bacterial growth, substituting the biological enrichment mechanism with a biochemical amplification mechanism that achieves similar sensitivity in hours rather than days.
Solution Approach 2:
The patent performs DNA extraction and PCR amplification as preliminary actions before detection. By preparing amplified DNA templates in advance through PCR, the system eliminates the need for time-consuming enrichment cultures while maintaining detection sensitivity, as the amplified DNA can be directly used for subsequent detection assays.
2Loss of time
If PCR testing is used to reduce detection time, then sensitivity and selectivity are improved, but the sample size that can be economically tested is limited
Solution Approach 1:
The patent segments the detection process into separate modules: DNA extraction, PCR amplification, and detection. This segmentation allows different sample volumes to be processed through the same protocol, enabling both small clinical samples and larger food/water samples to be tested economically by adjusting only the input sample volume rather than requiring different methodologies.
Solution Approach 2:
The patent creates a universal detection system where the same PCR-based methodology can be applied to various sample types and sizes. The system is designed to handle diverse samples (clinical, food, water) with different volumes using identical reagents and protocols, making the technique economically viable across a wide range of sample quantities.
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 substitutes the time-dependent biological enrichment process with a rapid PCR amplification process. Instead of relying on bacterial growth over overnight periods, the system uses enzymatic DNA amplification that achieves equivalent or superior sensitivity in 2-4 hours, enabling same-day results while maintaining high detection sensitivity.
Solution Approach 2:
The patent changes the fundamental parameter of time required for signal amplification by switching from biological growth (hours to days) to biochemical amplification (minutes to hours). By altering the amplification mechanism and its kinetic parameters, the system achieves high sensitivity without the time penalty of overnight enrichment, thereby improving productivity and turnaround speed.
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 achieves high sensitivity and specificity in detecting as few as 10 bacteria in a sample within a few hours, providing rapid and efficient detection of pathogens without the need for extensive culturing, suitable for food, water, and clinical samples.
Implementation Method 1
recombinant bacteriophages that are genetically modified to include a reporter gene, allowing for rapid detection of microorganisms by binding to them and generating an indicator moiety
Data Source
AI summary
Disclosed herein are devices, methods, and systems for rapid detection of microorganisms using a recombinant bacteriophage. The specificity of recombinant bacteriophages for binding microorganisms allows targeted and highly specific detection of a microorganism of interest.