Nucleotide Probe Detection of Active Toxic Algae at Low Cell Counts
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Solution Overview
Problem
Current methods for detecting toxic algae are lengthy, time-consuming, and have high detection limits, ranging from 1,000 to 12,500 cells/L, making them inadequate for rapid and reliable monitoring of toxic algal blooms in aquatic environments.
Innovation Solution
The use of specific nucleotide probes that target ribosomal nucleic acids (rRNA or rDNA) of active toxic algae, enabling detection and quantification at very low thresholds in less than one hour, with detection limits as low as 0.10 ng of RNA per liter and 100 to 500 active live cells per liter.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If sandwich hybridization is used for detecting toxic algae, then detection specificity is improved, but detection time increases and detection limit remains high (1,000-12,500 cells/L)
Solution Approach 1:
The detection system is segmented into multiple functional components: capture probes immobilized on solid support, signal probes with fluorescent labels, and specific oligonucleotide sequences targeting different toxic algal species. This segmentation allows parallel processing and rapid detection while maintaining specificity through targeted probe-algal nucleic acid interactions.
Solution Approach 2:
The patent replaces traditional mechanical microscopy examination with a molecular biology-based hybridization system using fluorescently labeled probes. This substitution enables automated, rapid detection with higher sensitivity, reducing detection time from hours to minutes while improving detection limits to 100-500 cells/L.
2Device complexity
If conventional detection methods are used, then equipment complexity is reduced, but detection sensitivity worsens (detection limit 1,000-12,500 cells/L)
Solution Approach 1:
Fluorescently labeled oligonucleotide probes serve as intermediaries between the detection system and target algal nucleic acids. These probes amplify the detection signal through fluorescent labeling, enabling highly sensitive detection of 100-500 cells/L without requiring complex instrumentation beyond standard fluorescence detection capabilities.
Solution Approach 2:
The patent changes the detection parameter from visual microscopy to fluorescent signal detection. By using fluorophore-labeled probes that emit detectable signals upon hybridization with target nucleic acids, the system achieves 10-100 times higher sensitivity while maintaining relatively simple equipment requirements through parameter transformation.
3Adaptability or versatility
If detection methods target all algal cells, then detection coverage is improved, but reliability worsens due to environmental variability and detection of non-toxic or dormant cells
Solution Approach 1:
The detection probes are designed with species-specific and toxin-specific local qualities through customized oligonucleotide sequences. Each probe targets specific genetic markers or toxin genes of particular toxic algal species, enabling selective detection that distinguishes toxic from non-toxic algae and active from dormant cells, thereby improving reliability while maintaining coverage of relevant targets.
Solution Approach 2:
The system performs preliminary enrichment and concentration of algal cells from water samples before detection. This preliminary action increases the proportion of detectable target cells while removing environmental inhibitors, improving both the reliability of toxin detection and the coverage of low-abundance toxic species in complex environmental matrices.
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 provides a rapid, sensitive, and reliable detection of active live cells of toxic algae, allowing for early warning of algal blooms and overcoming environmental variability, with detection limits significantly lower than existing methods.
Implementation Method 1
the use of specific probes and the implementation of the sandwich hybridization technique to specifically detect and quantify the nucleic acids of active live cells of toxic algae
Data Source
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AI summary
The invention relates to the use of probes to detect active living cells of toxic algae potentially present in a marine, brackish or industrial environment, a method for detecting active living cells of toxic algae potentially present in a marine environment, and corresponding kits.