Luminophore-Labeled Molecules for Microarray Assays
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Solution Overview
Problem
Current microarray-based assays require costly and inhibitory fluorescent labeling for target molecules, which complicates PCR reactions and increases costs, necessitating a more efficient and cost-effective labeling method for molecular detection.
Innovation Solution
A method involving the coupling of target molecules to particles with luminophores, where the luminophores are either directly or indirectly attached to the particles, allowing for efficient labeling of target molecules without interfering with PCR reactions, significantly reducing the required luminophore amount and costs.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescent labeling is used to label target molecules for detection, then detection sensitivity is improved, but cost increases and PCR reaction is inhibited
Solution Approach 1:
The patent segments the labeling process into two independent stages: (1) PCR amplification without labels, and (2) subsequent fluorescent labeling of amplified products. This separation allows PCR to proceed without inhibition, followed by efficient labeling of the already-amplified target molecules, thus resolving the contradiction between detection sensitivity and PCR inhibition
Solution Approach 2:
The patent performs PCR amplification as a preliminary action before fluorescent labeling. By first amplifying the target molecules without labels and then labeling the amplified products, the method eliminates PCR inhibition while maintaining detection sensitivity, as the labeling occurs after the critical amplification step is complete
2Measurement precision
If fluorescent labeling is used to label target molecules, then detection capability is improved, but cost increases
Solution Approach 1:
The patent applies partial labeling by using fluorescent labels to label only a subset of the amplified target molecules rather than all molecules. This partial action is sufficient for detection purposes while significantly reducing the total amount of expensive fluorescent labels required, thus lowering costs while maintaining detection capability
Solution Approach 2:
The patent uses the PCR amplification process to create multiple copies of the target sequence, and then labels these copies. By amplifying the target first and then labeling the copies, the method reduces the amount of expensive fluorescent label needed compared to labeling the original single-copy target molecules directly
3Measurement precision
If chemical modifications are made to primers for fluorescent labeling, then target molecules can be detected, but PCR reaction efficiency decreases
Solution Approach 1:
The patent segments the detection process into two distinct phases: (1) PCR amplification using unmodified primers for high efficiency, and (2) fluorescent labeling of the amplified products. This segmentation allows each process to optimize for its specific function without the negative effects of combining them
Solution Approach 2:
The patent performs PCR amplification as a preliminary action before fluorescent labeling. By completing the amplification first with unmodified primers and then adding fluorescent labels to the products, the method preserves PCR reaction efficiency while still achieving target detection through subsequent labeling
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 reduces the cost of fluorescence labeling and minimizes PCR inhibition, enabling high-sensitivity detection of molecular interactions with reduced luminophore usage, enhancing the efficiency and specificity of microarray-based assays.
Implementation Method 1
providing a luminophore on the target-particle complex, thereby directly or indirectly labeling the plurality of target molecules with the luminophore
Data Source
AI summary
A method for labeling target molecules coupled to particles for the detection of the target molecules using a microarray chip, comprises: providing a functionalized microparticle, wherein the microparticle is coated with one or more functional group; providing a modification group on each of the target molecules to be detected to form modified target molecules; contacting the functionalized microparticle with the modified target molecules; coupling a luminophore to the complex between the functionalized microparticle and the modified target molecules, thereby directly or indirectly labeling each modified target molecules with the luminophore. By directly or indirectly labeling the target molecules with the luminophore, the method reduces the cost of fluorescence detection, and avoids PCR inhibition derived from traditional fluorescence labeling molecules.