Multiplex Microorganism Detection via Fluorescent Probe Combinations
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting microorganisms in samples are laborious and slow, necessitating the development of rapid, sensitive, and accurate techniques for identifying multiple microorganisms simultaneously.
Innovation Solution
The use of probes labeled with combinations of detectable labels allows for the detection of multiple microorganisms using a minimum number of probes, leveraging unique label combinations for each microorganism, enabling a multiplex assay that can identify a wide range of microorganisms efficiently.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional culture methods are used to detect microorganisms, then identification accuracy is maintained, but detection time and labor requirements increase significantly
Solution Approach 1:
The patent replaces traditional mechanical/culturing methods with molecular biological techniques. Specifically, it uses fluorescent in situ hybridization (FISH) with fluorescently labeled probes that bind to ribosomal RNA of microorganisms, enabling direct detection without culturing. This substitution of detection mechanism dramatically reduces detection time while maintaining identification accuracy through sequence-specific probe binding.
Solution Approach 2:
The patent changes the detection parameter from growth-based identification to nucleic acid hybridization. By detecting fluorescent signals from probes bound to ribosomal RNA sequences, the method transforms the detection paradigm from requiring microbial growth (time-consuming) to direct molecular detection (rapid), thereby reducing detection time while preserving accuracy through sequence-specific targeting.
2Adaptability or versatility
If multiple microorganisms are detected using separate probe assays, then detection comprehensiveness improves, but assay complexity and reagent requirements increase
Solution Approach 1:
The patent creates a universal detection system where a single fluorescent probe can detect multiple different microorganisms. The probe is designed with a universal fluorescent label that can be detected by the same fluorescence microscope, allowing one assay to simultaneously identify various bacterial species, fungi, and other microorganisms present in the sample, thereby achieving multi-functionality without proportionally increasing complexity.
Solution Approach 2:
The patent merges multiple detection functions into a single assay by using fluorescent probes that can be visualized together on the same microscope slide. Instead of requiring separate assays for different microorganisms, the method combines detection of multiple targets into one unified fluorescent microscopy experiment, reducing overall assay complexity while maintaining comprehensive detection capability.
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 straightforward detection of multiple microorganisms, providing timely information for selecting appropriate therapeutic regimens and reducing the need for extensive culturing processes.
Implementation Method 1
contacting the sample with a plurality of detectably-labeled, microorganism-specific probes... under conditions that allow hybridization of the probes to a nucleic acid sequence of the first microorganism
Implementation Method 2
probes labeled with combinations of detectable labels... The first and second labels are selected such that the first detectable label provides a first signal, the second detectable label provides a second signal
Data Source
AI summary
Presented herein are methods for the detection of the presence or absence of one or more microorganisms in a sample. The method deploys a plurality of probe sets to detect a plurality of microorganisms. The probes in the probe set are detectably labeled. At least one probe set has probes labeled with a combination of detectable labels. The number of detectable labels used in the plurality of probe sets numbers less than the number of microorganisms being detected by the probe set.