Nuclease-Activity Composition for On-Site Microbial Detection
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Solution Overview
Problem
Existing methods for detecting microbial contamination are time-consuming, require expensive equipment, and produce false positives due to the persistence of ATP signals from dead or low-viability microorganisms, making rapid and accurate detection challenging.
Innovation Solution
A solid substrate-immobilized oligonucleotide probe linked to magnetic nanoparticles, which detects microbial contamination by measuring the activity of nuclease enzymes using a label that generates a detectable signal upon cleavage by microbial nucleases.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If ATP measurement is used to detect microorganisms on-site, then detection speed is improved, but measurement accuracy deteriorates due to false positives from dead or low-viability microorganisms
Solution Approach 1:
The patent changes the detection parameter from ATP presence (which persists in dead cells) to nuclease activity measurement. By using a nuclease-specific inhibitor and detecting the inhibition effect, the method specifically targets living microorganisms that possess active nucleases, thereby eliminating false positives from dead cells while maintaining rapid detection capability
Solution Approach 2:
The patent introduces a nuclease-specific inhibitor as an intermediary substance. This inhibitor specifically binds to active nucleases in living microorganisms, creating a detectable signal that distinguishes viable cells from dead ones. The inhibitor acts as a mediator that converts the biological activity of nucleases into a measurable detection signal
2Measurement precision
If PCR or ELISA methods are used to detect pathogens, then measurement precision is improved, but device complexity and operation difficulty increase due to expensive equipment and skilled technology requirements
Solution Approach 1:
The patent extracts the core detection function from complex laboratory equipment and simplifies it to a biochemical assay that can be performed with basic laboratory tools. By focusing solely on nuclease activity detection using simple inhibition-based methodology, the patent removes the need for expensive PCR machines or ELISA equipment while maintaining detection accuracy
Solution Approach 2:
The patent employs simple, inexpensive reagents including nuclease-specific inhibitors and basic detection substrates that can be used without sophisticated equipment. The method relies on straightforward biochemical reactions that can be performed in standard laboratory settings, replacing expensive and complex diagnostic tools
3Measurement precision
If conventional culture methods are used to detect microorganisms, then measurement precision is improved, but loss of time increases due to long proliferation and culture periods
Solution Approach 1:
The patent performs preliminary detection of nuclease activity directly from environmental samples without requiring microorganisms to be cultured first. By detecting the enzymatic activity of nucleases that are already present in viable cells, the method eliminates the time-consuming culture proliferation step while maintaining the ability to accurately identify pathogenic microorganisms
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 and accurate detection of living microorganisms, eliminating false positives and requiring no specialized equipment, suitable for various bacteria and fungi.
Implementation Method 1
a solid surface-immobilized probe having one end bound to a solid substrate and the other end bound to an enzyme capable of exhibiting a signal to measure the activity of a nuclease present in cells
Implementation Method 2
measure the activity of enzymes always present in cells to apply to detection of microorganisms
Data Source
AI summary
The present invention relates to a composition for detecting on-site microbial contamination and uses thereof. The composition for detecting on-site microbial contamination, according to the present invention, is capable of detecting the nucleic acid decomposition ability of a comprehensive nuclease, and detecting a living microorganism in a sample very quickly and accurately, and a detection method thereby is very simple. Accordingly, the composition of the present invention is expected to be able to easily and conveniently measure the degree of contamination of microorganisms in an environment.