Microorganism Detection via Ribosome Extraction and Phage Amplification
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Solution Overview
Problem
Current methods for detecting bacteria and other microorganisms in samples are time-consuming, often requiring several days due to the need for biological enrichment and cultivation, which is unsuitable for rapid identification of pathogens in food, water, or clinical samples, especially with the rise of antibiotic-resistant bacteria and biodefense concerns.
Innovation Solution
The use of ribosomes and bacteriophage-based methods for amplifying detection signals, where ribosomes or progeny bacteriophage are isolated and detected using bead-based amplified immunoassays or lateral flow assays, allowing for rapid and sensitive detection of microorganisms without the need for traditional enrichment cultures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional microbiological tests with enrichment cultures are used, then sensitivity of detection is improved, but detection time increases to several days
Solution Approach 1:
The patent extracts ribosomes from bacterial cells using lysis and separates them from cell debris through filtration and washing steps. This extraction allows detection of the bacterial signature (ribosomes) without requiring the time-consuming growth of entire bacterial cultures, thereby reducing detection time while maintaining sensitivity
Solution Approach 2:
The patent uses ribosomes as an intermediary marker to detect bacterial presence. Instead of detecting whole bacteria directly which requires enrichment, the method detects ribosomal RNA or proteins released from lysed cells. This intermediary approach enables sensitive detection without the need for prolonged cultural enrichment
2Measurement precision
If PCR amplification is used to achieve high sensitivity, then detection sensitivity is improved, but the sample size that can be economically tested is limited
Solution Approach 1:
The patent creates multiple copies of the detection signal by lysing each bacterial cell to release thousands of ribosomes, which can then be detected. This natural copying mechanism within each cell provides signal amplification similar to PCR but without the need for expensive thermal cyclers, enabling higher throughput testing
Solution Approach 2:
The patent replaces the complex mechanical and thermal system of PCR amplification with a simpler lysis-based release system. By mechanically or chemically lysing cells to release ribosomes, the method achieves sensitivity without requiring the sophisticated equipment and protocols of PCR, thereby increasing accessibility and throughput
3Loss of time
If direct immunoassays are used for rapid detection, then detection time is reduced, but sensitivity decreases requiring enrichment steps
Solution Approach 1:
The patent performs preliminary lysis of bacterial cells to release ribosomes before the immunoassay detection step. This preliminary action concentrates the detectable target (ribosomes) from a small volume of lysate, providing sufficient signal for sensitive detection without requiring prior enrichment of whole bacteria
Solution Approach 2:
The patent changes the physical state of bacterial components from intact cells to released ribosomal fragments through lysis. This parameter change transforms the target from whole cells requiring enrichment to abundant intracellular fragments that provide strong signal in direct immunoassays, achieving both rapid detection and high 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 the detection of microorganisms, including single bacterial cells, in a short time frame, providing high sensitivity and specificity, and overcoming the limitations of traditional methods by amplifying signals through ribosome or bacteriophage detection, facilitating rapid identification of pathogens.
Implementation Method 1
The ribosomes and/or ribosomal proteins released from the microorganism may be assayed using a bead-based amplified immunoassay
Implementation Method 2
the present invention may comprise a lateral flow assay in combination with carbon black nanostrings to detect ribosomes and/or ribosomal proteins released from the microorganism
Implementation Method 3
utilizes the high specificity of agents that can bind to microorganisms or their constituents as a means to detect and isolate low levels of a microorganism
Data Source
AI summary
Disclosed are methods and systems for the isolation and detection of microbes from a sample. The use of binding agents for isolation of a microbe of interest from a sample are described. In certain embodiments, the methods use ribosome-based and/or bacteriophage-based amplification of the signal in detection of bacteria and other microorganisms. For example, embodiments of the present invention can achieve total amplification of at least 10,000 from a single infected cell.