Fluorescence Assay for Pathogen Identification
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting pathogens in bodily fluids are slow, costly, and limited in sensitivity and specificity, often requiring complex sample preparation and specific reagents, making them unsuitable for rapid and broad pathogen identification, especially for new or unknown types.
Innovation Solution
A fluorescence microscopy method using non-specific fluorescent probes that bind to pathogen surface carbohydrates, nucleic acid, and membrane components, generating a distinctive 'signature' for identification, which can be applied to various pathogens with minimal sample preparation and without the need for specific antibodies or nucleic acid sequences.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If conventional microscopy-based detection methods are used, then the method is simple and quick, but the sensitivity is low and can only detect large pathogens with high concentration
Solution Approach 1:
The patent applies fluorescence labeling to pathogens, transforming them from non-fluorescent to fluorescent states. Different pathogens are labeled with different fluorescent colors (e.g., green for bacteria, red for viruses), enabling highly sensitive detection through fluorescence microscopy while maintaining rapid detection speed. This color-based differentiation resolves the contradiction between simple quick detection and sensitive detection.
2Measurement precision
If fluorescence-based techniques with specific binding molecules are used, then the identification accuracy for specific pathogen types is improved, but the cost increases and it becomes impractical for broad spectrum testing
Solution Approach 1:
The patent uses universal fluorescent labeling reagents (such as fluorescent dyes that bind to common pathogen components like carbohydrates, nucleic acids, or membranes) that can label multiple types of pathogens simultaneously. This multi-functional approach allows a single reagent system to detect broad spectrum of pathogens while maintaining identification accuracy through fluorescence microscopy, avoiding the need for multiple specific antibodies or aptamers.
3Measurement precision
If established methods like PCR or antigen detection are used, then the detection accuracy is high, but the test time increases to hours and the cost increases due to expensive reagents
Solution Approach 1:
The patent replaces complex biochemical amplification systems (PCR enzymes, antigen-antibody reactions) with direct fluorescence labeling and optical detection. This substitution eliminates the need for time-consuming enzymatic reactions and amplification steps, reducing test duration from hours to minutes while maintaining high detection accuracy through the sensitivity of fluorescence microscopy.
4Measurement precision
If electron microscopy is used, then the detection sensitivity for small pathogens is improved, but the sample preparation complexity and time increase significantly
Solution Approach 1:
The patent introduces fluorescent labels as intermediaries that bind to pathogens and provide optical contrast visible under fluorescence microscopy. This intermediary approach allows detection of small pathogens without requiring the complex vacuum-based imaging and extensive sample preparation of electron microscopy, as the fluorescent labels amplify the optical signal of small pathogens making them detectable by simpler, faster fluorescence microscopy methods.
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, cost-effective, and accurate identification of pathogens in bodily fluids, including new types, with minimal sample preparation, and can be adapted for pre-emptive mass testing and screening without prior knowledge of virus genetics or biology.
Implementation Method 1
A fluorescence microscopy method using non-specific fluorescent probes that bind to pathogen surface carbohydrates, nucleic acid, and membrane components, generating a distinctive 'signature' for identification
Data Source
AI summary
The present application discloses a method of identifying and characterising intact pathogens in a sample using a fluorescence imaging system, in particular for characterising bacteria and viruses, and associated systems and kits for implementing the imethod. The method involves incubating a sample with fluorescent probes from at least two of the following categories: a fluorescent probe for binding to a pathogen surface carbohydrate; a nucleic acid stain; and a membrane stain. The sample is then imaged using fluorescence imaging apparatus to detect candidate objects, and the fluorescence characteristics of the candidate objects used to identify whether the object is a pathogen and, if so, the type of pathogen. Particularly preferred implementations employ flow to improve data acquisition, and machine learning classification algorithms to distinguish different pathogen types.