Microbial Detection via Label Molecule Complex Formation
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Solution Overview
Problem
Conventional microbial testing methods are costly and time-consuming, especially when samples contain no microorganisms, and require complex procedures involving enzyme use and purification steps.
Innovation Solution
A method involving the formation of a complex using label molecules A, B, and C, which hybridize with nucleic acid targets, allowing for rapid microbial inclusion determination and identification without the need for expensive reagents or enzymes, utilizing phosphor-labeled molecules and magnetic or metal microparticles for enhanced detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional microbial testing procedures are performed on all samples including those with no microorganisms, then microbial inclusion can be determined, but testing cost increases significantly
Solution Approach 1:
The patent applies partial action by performing amplification only on samples that show positive results in the initial screening step. Instead of subjecting all samples to the complete 4-step procedure, only samples with detected amplification signals proceed to microarray detection, thereby reducing reagent consumption and testing costs while maintaining reliable microbial inclusion determination.
2Measurement precision
If real-time PCR is performed to determine bacterial presence, then amplification results can be obtained, but the procedure becomes complicated and requires purification steps
Solution Approach 1:
The patent extracts and eliminates the purification step from the conventional 4-step procedure. By using a simplified amplification approach where the amplification products are directly applied to microarray detection without intermediate purification, the method maintains measurement precision while significantly reducing procedural complexity and eliminating the need for additional reagents and equipment.
Solution Approach 2:
The patent merges the amplification and detection steps by directly applying amplification products to microarray detection. This consolidation eliminates the separate purification step, reducing overall procedure complexity while maintaining the ability to obtain accurate amplification results for microbial detection.
3Measurement precision
If enzyme-based amplification is used to detect microorganisms, then detection sensitivity is improved, but testing cost increases due to enzyme reagents
Solution Approach 1:
The patent replaces expensive enzyme-based reagents with cost-effective oligonucleotide primers and probes for amplification and detection. The method uses simple DNA polymerase or other non-enzyme amplification techniques combined with microarray detection, achieving comparable detection sensitivity while dramatically reducing reagent costs by eliminating the need for expensive enzyme-based kits.
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 method simplifies microbial testing by reducing costs and time, enabling efficient detection and identification of microorganisms with high brightness in fluorescence observation, even at low concentrations, without the need for enzyme-based purification.
Implementation Method 1
the label molecule A comprises a nucleic acid sequence capable of hybridizing to a part of the target and substantially complementary to the nucleic acid sequence of the target
Implementation Method 2
at least one of either the label molecule A or the label molecule C has a phosphor
Implementation Method 3
the label molecule B has at least one of either magnetic microparticles or metal microparticles
Data Source
AI summary
A method for producing a complex, a method for determining microbial inclusion, and a method for identifying the included microorganism, which can be easily performed is provided. A method for producing a complex containing a target that is contained in a sample, includes a step (a) of bringing a sample into contact with a label molecule A, a label molecule B, and a label molecule C and forming a complex, and a step (b) of isolating the complex. The target is a nucleic acid. The label molecule A contains a nucleic acid sequence capable of hybridizing to a part of the target and substantially complementary to the nucleic acid sequence of the target. The label molecule B contains a nucleic acid sequence capable of hybridizing to a part of the target and substantially complementary to the nucleic acid sequence of the target.