Digital Counting via Repeated Signal Deactivation
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Solution Overview
Problem
Current digital counting methods face challenges in reducing false-positive detections and background noise, particularly in complex samples like blood, due to non-specific binding and high concentrations of non-target molecules, which decrease sensitivity and specificity.
Innovation Solution
The use of a plurality of discrete capture sites for analytes, with multiple detection cycles that involve labeling and deactivation of signals, reduces non-specific interactions and enhances specific binding, thereby minimizing counting errors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple detection cycles with labeling and deactivation are used, then false-positive detections and background noise are reduced, but the complexity of the detection process increases
Solution Approach 1:
The patent implements periodic detection cycles where the same capture sites undergo repeated labeling and signal deactivation steps. This periodic action allows true positive signals to be consistently detected across multiple cycles while false positives (which do not consistently produce signals) can be statistically identified and eliminated, thereby improving measurement precision through iterative verification
Solution Approach 2:
The patent performs preliminary deactivation of signals between detection cycles to eliminate false positives before the next labeling step. By preemptively removing non-specific signals and resetting the system state, the method prepares the capture sites for the next detection cycle in a controlled manner, ensuring that only consistent true positive signals accumulate across cycles
2Measurement precision
If multiple detection cycles are performed, then counting errors are minimized, but the time required for analysis increases
Solution Approach 1:
The patent employs periodic detection cycles with standardized labeling and deactivation steps that can be efficiently repeated. By organizing the detection process into discrete, reusable cycles, the method minimizes the time overhead between cycles while ensuring that counting accuracy improves through the accumulation of consistent positive signals across multiple iterations
Solution Approach 2:
The patent maintains continuous useful action by immediately transitioning from signal deactivation to the next labeling step without unnecessary idle time. The deactivation step prepares the system for the next detection cycle, ensuring that the time between cycles is productively used rather than lost, thereby minimizing total analysis time while still achieving improved counting accuracy through multiple cycles
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 significantly reduces false-positive detections and background noise, improving the sensitivity and specificity of digital counting analyses by ensuring consistent signal production from target analytes while suppressing non-specific interactions.
Implementation Method 1
single immunocomplexes comprising an analyte molecule sandwiched between a capture antibody and an enzyme-linked detection antibody
Implementation Method 2
supplied with fluorogenic or chromogenic enzyme substrate to produce a detectable optical signal within the compartment
Implementation Method 3
single oligonucleotide analytes are co-encapsulated with PCR primers and PCR mixture in compartments, hence leading to the analyte-templated exponential amplification of fluorescence-labeled amplicons
Data Source
AI summary
The present invention relates to methods and systems for testing for the presence of a material such as one or more analyte types within a sample and more particularly, for improved single enzyme-linked immunosorbent assay (sELISA) testing as well as other variants of single-enzyme linked molecular analysis (SELMA). Background and false positives are reduced due to the presence of at least two detection cycles where each detection cycle comprises the steps of a) triggering a signal from captured and labelled analyte(s), b) recording of the number and positions of capture sites exhibiting a signal from the captured and labelled analyte(s), c) and before a further detection cycle is performed, deactivation of signal(s).