Single-Step Analyte Detection Using Identical Hybridization Probes
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Solution Overview
Problem
Current diagnostic assays face challenges in efficiently identifying analytes in multiplex reactions without spatial separation, requiring multiple labels and re-testing to differentiate between multiple analytes, which increases complexity and time, especially as the number of analytes increases.
Innovation Solution
A method and apparatus that use a single read system with identical or distinguishable detectable labels for internal control and analyte probes, measuring combined signals without distinguishing between them, and employing multiple thresholds to determine analyte presence, allowing for simultaneous detection of multiple analytes in a single reaction mixture at constant temperature.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If multiple labels are used to detect multiple analytes in a multiplex reaction without spatial separation, then the ability to detect multiple analytes simultaneously is improved, but the complexity of the assay and the need for re-testing increases
Solution Approach 1:
The patent segments the detection of multiple analytes by using multiple probes with identical labels in separate detection channels. Each probe targets a specific analyte, and the signals are differentiated through computational analysis rather than physical separation, allowing simultaneous detection while managing complexity through software-based discrimination.
Solution Approach 2:
The patent applies universality by using identical detectable labels on all probes (both analyte and internal control probes) so that a single label type can serve multiple detection functions. This universal labeling approach simplifies the assay design compared to using different labels for different analytes, reducing the need for multiple reagent sets while maintaining the ability to detect multiple targets.
2Adaptability or versatility
If multiple labels are used to distinguish between multiple analytes in a multiplex reaction, then the detection capability is improved, but the time required for re-testing increases
Solution Approach 1:
The patent performs preliminary action by incorporating an internal control probe with identical labeling into the multiplex reaction mixture before detection. This internal control serves as a reference that is established in advance, allowing the system to distinguish between analyte signals and background noise without requiring subsequent re-testing or time-consuming verification steps.
Solution Approach 2:
The patent uses feedback through the internal control probe, which provides a reference signal that feeds back into the analysis. By comparing the signals from multiple analyte probes against the internal control signal, the system can automatically determine positive results without requiring re-testing, thereby reducing time loss while maintaining high detection capability.
3Ease of operation
If spatial separation is used to distinguish between internal control and analyte probes, then the ease of detection is improved, but the device complexity increases
Solution Approach 1:
The patent replaces the mechanical/spatial separation system with a computational/digital system. Instead of physically separating internal control and analyte probes in different spatial locations, the patent uses identical labels for both and relies on computational algorithms to distinguish their signals. This substitution eliminates the need for complex spatial separation mechanisms while maintaining ease of detection through automated signal analysis.
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 simplifies analyte identification by reducing the need for spatial separation and multiple labels, enabling efficient detection of multiple analytes in a single read, thereby reducing the burden of re-testing and improving assay efficiency.
Implementation Method 1
nucleic acid amplification followed by probe hybridization and detection
Implementation Method 2
an optical detection mechanism arranged to receive optical signals from the sample
Data Source
AI summary
Kits for detecting analyte polynucleotides and an internal control in a sample. Included in the kit are an internal control polynucleotide and amplification reagents to co-amplify a first analyte polynucleotide and the internal control. Also included are first and second hybridization probes, each having a label indistinguishable from the other. The probes are respectively capable of hybridizing with a first analyte amplicon and an internal control amplicon. The first and second labels are indistinguishable homogeneous labels.