Resolving Agents for Multiplex Nucleic Acid Detection

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Solution Overview

Problem

Existing nucleic acid detection methods face challenges in resolving differentially labelled rolling circle amplification (RCA) products in multiplex reactions due to label interference, which affects the accuracy and sensitivity of target nucleic acid sequence detection.

Innovation Solution

Incorporating resolving agents, such as proteins or detergents, into the reaction mixture to inhibit label interaction between different nucleic acid products, thereby improving the resolution of differently labelled RCA products.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If multiple differently labelled RCA products are detected in a multiplex reaction mixture, then the ability to detect multiple target nucleic acid sequences simultaneously is improved, but label interference between different nucleic acid products worsens the resolution and accuracy of detection

Engineering Contradiction:

Improvemultiplex detection capabilityVSAvoidresolution of differently labelled products

Core Design Contradiction:

Adaptability or versatilityVSMeasurement precision

Solution Approach 1:

The patent introduces resolving agents (such as proteins, detergents, or other molecules) as intermediary substances that interact with the labels on different RCA products to reduce or eliminate label interference. These resolving agents mediate the detection process by preventing unwanted interactions between labels from different nucleic acid products, thereby improving resolution while maintaining multiplex detection capability

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent modifies the chemical or physical parameters of the detection system by adding resolving agents that change the interaction properties between labels. This parameter change (introducing new molecular interactions) allows multiple labelled products to coexist in the same reaction mixture without interference, enabling both high multiplex capability and accurate resolution

Inventive Principle:

Principle #35Parameter changes

2Reliability

If the amount of labelled nucleic acid products is increased to improve detection sensitivity, then the sensitivity of target nucleic acid detection is improved, but label interference between products worsens the resolution

Engineering Contradiction:

Improvedetection sensitivityVSAvoidresolution of labelled products

Core Design Contradiction:

ReliabilityVSMeasurement precision

Solution Approach 1:

Resolving agents serve as intermediary molecules that prevent direct interference between labels even when multiple labelled RCA products are present in high concentrations. This allows the system to maintain both high sensitivity (through increased label amounts) and high resolution (through mediator prevention of label interactions)

Inventive Principle:

Principle #24Intermediary (Mediator)

3Use of energy by moving object

If more labels are attached to each nucleic acid product to enhance signal strength, then the detection signal is improved, but the interference between labels from different products worsens the resolution

Engineering Contradiction:

Improvesignal strengthVSAvoidresolution of differently labelled products

Core Design Contradiction:

Use of energy by moving objectVSMeasurement precision

Solution Approach 1:

The patent converts the potentially harmful effect of label interference into a beneficial situation by using resolving agents that specifically target and prevent label-label interactions. The high label density that would normally cause interference is instead harnessed to provide strong signals, while the resolving agents ensure that the interference is eliminated, making both high signal strength and high resolution achievable simultaneously

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

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 use of resolving agents enhances the ability to distinguish and detect multiple nucleic acid sequences in a single reaction mixture, improving the accuracy and sensitivity of nucleic acid detection assays.

Implementation Method 1

Incorporating resolving agents, such as proteins or detergents, into the reaction mixture to inhibit label interaction between different nucleic acid products

Methodology Applied

Scientific EffectLabel interaction inhibition:

Data Source

PatentUS20250179560A1Method for detecting nucleic acid amplification products using blocking agents

Publication Date: 2025.06.05 RARITY BIOSCIENCE AB

AI summary

The present invention provides a method for detecting two or more differently labelled high molecular weight nucleic acid products in a mixture, wherein said products are detected by detecting one or more labels which are attached directly or indirectly to each product, said method comprising including in the mixture for detection a resolving agent to facilitate resolution of the differently labelled products, wherein said resolving agent acts to inhibit interaction between different labels.